Subcloning – Wikipedia

In molecular biology, subcloning is a technique used to move a particular DNA sequence from a parent vector to a destination vector.

Subcloning is not to be confused with molecular cloning, a related technique.

Restriction enzymes are used to excise the gene of interest (the insert) from the parent. The insert is purified in order to isolate it from other DNA molecules. A common purification method is gel isolation. The number of copies of the gene is then amplified using polymerase chain reaction (PCR).

Simultaneously, the same restriction enzymes are used to digest (cut) the destination. The idea behind using the same restriction enzymes is to create complementary sticky ends, which will facilitate ligation later on. A phosphatase, commonly calf-intestinal alkaline phosphatase (CIAP), is also added to prevent self-ligation of the destination vector. The digested destination vector is isolated/purified.

The insert and the destination vector are then mixed together with DNA ligase. A typical molar ratio of insert genes to destination vectors is 3:1;[1] by increasing the insert concentration, self-ligation is further decreased. After letting the reaction mixture sit for a set amount of time at a specific temperature (dependent upon the size of the strands being ligated; for more information see DNA ligase), the insert should become successfully incorporated into the destination plasmid.

The plasmid is often transformed into a bacterium like E. coli. Ideally when the bacterium divides the plasmid should also be replicated. In the best case scenario, each bacterial cell should have several copies of the plasmid. After a good number of bacterial colonies have grown, they can be miniprepped to harvest the plasmid DNA.

In order to ensure growth of only transformed bacteria (which carry the desired plasmids to be harvested), a marker gene is used in the destination vector for selection. Typical marker genes are for antibiotic resistance or nutrient biosynthesis. So, for example, the "marker gene" could be for resistance to the antibiotic ampicillin. If the bacteria that were supposed to pick up the desired plasmid had picked up the desired gene then they would also contain the "marker gene". Now the bacteria that picked up the plasmid would be able to grow in ampicillin whereas the bacteria that did not pick up the desired plasmid would still be vulnerable to destruction by the ampicillin. Therefore, successfully transformed bacteria would be "selected."

In this example, a gene from mammalian gene library will be subcloned into a bacterial plasmid (destination platform). The bacterial plasmid is a piece of circular DNA which contains regulatory elements allowing for the bacteria to produce a gene product (gene expression) if it is placed in the correct place in the plasmid. The production site is flanked by two restriction enzyme cutting sites "A" and "B" with incompatible sticky ends.

The mammalian DNA does not come with these restriction sites, so they are built in by overlap extension PCR. The primers are designed to put the restriction sites carefully, so that the coding of the protein is in-frame, and a minimum of extra amino acids is implanted on either side of the protein.

Both the PCR product containing the mammalian gene with the new restriction sites and the destination plasmid are subjected to restriction digestion, and the digest products are purified by gel electrophoresis.

The digest products, now containing compatible sticky ends with each other (but incompatible sticky ends with themselves) are subjected to ligation, creating a new plasmid which contains the background elements of the original plasmid with a different insert.

The plasmid is transformed into bacteria and the identity of the insert is confirmed by DNA sequencing.

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Subcloning - Wikipedia

Card cloning racket: Waiters earned 1000 for each card they sold details of, say Mumbai cops – Hindustan Times

The investigation into a card cloning racket, unearthed by the Bandra police this month, has revealed that the racketeers allegedly paid Rs1,000 to each waiter for sharing details,including password, of each debit or credit card of customers.

The police last week arrested six waiters who worked at restaurants in Mumbai, Pune and Thane and two ITexperts. The waiter allegedly stole credit and debit card details of 1,028 customers. Officials said IT professionals had contacted the waiters through an acquaintance in the hotel management industry.

Pandit Thackeray, senior inspector of Bandra police station, confirmed that each waiter was paid Rs1,000 for each card.

By selling credit and debit card details, each waiter earned around Rs50,000 per month, according to an officer.

Thackeray added that the duped customers had accounts in 108 banks. The police have recovered 106 skimmers devices used to obtain card details from the eight men. The accused men made these devices from the scratch and uploaded the stolen data on empty cards. The accused then withdrew money from the customers account using these cloned cards. The waiters in the beginning copied details of one or two card. As they grew confident, they began cloning 40 to 50 cards in a month, added the police officer.

Three of these waiters worked at a restaurant at Sakinaka in Andheri (East). The unsuspecting customers handed over their cards and pin code to the waiters who secretly swiped them on the skimmers. In some cases, they clandestinely saw customers punching in card pin code on the swipe machine.

The Bandra police received a complaint from Citi Bank in May which led to the racket. Apart from the 106 skimmers, the police seized a laptop and a unique software used to clone cards. The police have also seized 50 cloned cards from the accused which are rewritable.

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Card cloning racket: Waiters earned 1000 for each card they sold details of, say Mumbai cops - Hindustan Times

Organ Rejection’s Innate Instigator Cornered by Positional Cloning – Genetic Engineering & Biotechnology News (blog)

In hopes of heading off transplantation rejection before it can even get started, scientists based at the University of Pittsburgh and the University of Toronto have been mapping the molecular landscape of the innate immune response. This territory is relatively unfamiliar, compared to the adaptive immune response, yet it needs to be explored because it is where the very first steps in rejection pathways may be found.

Curiously, when the adaptive immune system is deliberately compromised in experimental mice, the innate immune system, which remains intact, can still detect allogeneic transplants and instigate the first step of a rejection process, specifically, the generation of mature dendritic cells. Ordinarily, these cells, which produce interleukin-12, ultimately present antigen to T cells. How this step in the rejection process is instigated hasnt been clear. That is, its molecular mechanisms havent been elucidated.

Reasoning that these molecular mechanisms might be relevant to humans as well as mice, the Pittsburgh/Toronto scientists decided to hunt them down. To keep the hunt on track, the scientists relied on a classical genetic mapping technique called positional cloning, which narrows down a candidate region of the genome, thought to be involved in some phenotype, to a candidate gene.

Details of the Pittburgh/Tornonto teams genetic pursuit appeared June 23 in the journal Science Immunology, in an article entitled Donor SIRP Polymorphism Modulates the Innate Immune Response to Allogeneic Grafts. It describes how a positional cloning approach was used to identify polymorphisms in the mouse gene encoding signal regulatory protein (SIRP). It goes on to assert that these polymorphisms are key to innate self-nonself recognition.

We studied the innate response of Rag2/c/ mice, which lack T, B, and NK [natural killer] cells, to grafts from allogeneic donors, wrote the articles authors, who added that they identified the donor polymorphism in SIRP as a key modulator of the recipients innate allorecognition response. Donors that differed from the recipient in one or both Sirpa alleles, they explained, elicited an innate alloresponse. The response was mediated by binding of donor SIRP to recipient CD47 and was modulated by the strength of the SIRP-CD47 interaction.

Essentially, when the transplanted tissue SIRP is different from the host tissue SIRP, the transplant SIRP binds to CD47, which is located on the recipient's monocytes, a class of innate immune cells. This binding kicks off a series of cellular events that activate the innate and then eventually the adaptive immune system.

"For the first time, we have an insight into the earliest steps that start the rejection response," asserted the study's co-senior author Fadi Lakkis, M.D., Frank & Athena Sarris Chair in Transplantation Biology and scientific director of Pitt's Thomas E. Starzl Transplantation Institute. "Interrupting this first recognition of foreign tissues by the innate immune system would disrupt the rejection process at its earliest inception stage and could prevent the transplant from failing."

Approximately 50% of all transplanted organs are rejected within 10 to 12 years, so there is a great need for better ways to reduce or eliminate organ rejection.

Like mice, humans also express SIRP, so sequencing the gene to identify donors and recipients with matched forms of the molecule hopefully will lead to lower organ rejection rates in the future, Lakkis said.

Blocking the interaction between SIRP and CD47 in mice prevented the monocyte activation, suggesting that disruption of this coupling could prevent recipient immune system activation. Future studies to examine how the interaction between SIRP and CD47 leads to monocyte activation could lead to new ways to prevent organ rejection.

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Organ Rejection's Innate Instigator Cornered by Positional Cloning - Genetic Engineering & Biotechnology News (blog)

Fraud Expert: Credit Card Cloning Trend "Going Skyward" – WFYI

Photo by Flickr user Sean MacEntee

A recent weekend getaway with the family turned into a credit card cloning hassle for a Pendleton man. Luke Renner had taken his family to an amusement park, only to discover a few days later that two of his credit cards had been cloned.

My best guess that it probably - whatever happened happened there," Renner says.

Renner received a fraud alert from one of the banks to let him know that his personal credit card had been hacked. It was only after his business credit card was declined at two different stores that he realized it too had been cloned.

And then it occurred to me that one of the two cards that was compromised never left my wallet, he says.

Renner didn't take any chances. He and his wife closed their credit card accounts and got new cards. They also did some research on RFID or so-called "tap-to-pay" credit cards that you don't have to swipe or stick in a machine to use.

They have RFID blocking wallets, card sleeves that stick to your phone and these are just sort of barriers that create an electronic wall of some sort that keep those scanners from being able to retrieve the information off of the cards that are in your wallet, Renner says.

He's invested in a new wallet for himself and his wife got an RFID blocking sleeve for her cards.

Indianapolis Metro Police Sergeant Steve Walters says credit card fraud continues to escalate. Criminals use skimmers on ATMs and at gas pumps but, he says, they can also scan tap-to-pay credit cards without even touching them.

They can do a card reading right from the pocket of their pants and you have no idea. It could have been someone standing right behind you at the grocery store, Walters says.

He recommends setting up fraud alerts on all your credit cards and regularly - even daily - check your statements online.

Walters warns there may be more to worry about than just getting your credit card replaced.

Make sure in the next couple of weeks once you have had that account shut down, that you run a credit check on yourself to make sure individuals are not opening different accounts using your card information, he says.

Walters also recommends making a police report so they can help put a stop to credit card fraud.

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Fraud Expert: Credit Card Cloning Trend "Going Skyward" - WFYI

Youth’s arrest unearthed card cloning fraud – Times of India

MUMBAI: The arrest of a 30-year-old college dropout from Byculla had helped the police unearth a major card cloning racket and crack 14 cases three in Bandra and the rest in Thane and Pune. Till date, eight men have been arrested and two kignpins are on the run. Police said the rackeeters, with the help of six hotel waite-rs, stole debit/credit card details of 1,000 people from 96 banks, cloned cards and withdrew money from unmanned ATM kiosks. On June 18 night, police sub-inspector Bhimsen Gaikwad noticed a suspicious-looking man inside an ATM kiosk on Carter Road in Bandra (W). The man, Sayyed Masrafe, claimed he was trying to withdraw money, but Gaikwad found it was an expired card. "During questioning, we learnt that he was on bail after being arrested in another card fraud case," said a police officer. "We searched Masrafe's house and found a cloning app on his laptop, skimmers, internet dongle, a coding and decoding machine to make magnetic strips and 50 forged debit cards. The arrested waiters were given skimmers to swipe customers' cards, while Masrafe would clone the cards and withdraw money from ATMs."

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Youth's arrest unearthed card cloning fraud - Times of India

Card cloning racket: Mumbai gang steals debit, credit card data from 1000 customers of 96 banks – Hindustan Times

Investigations into a card cloning racket which was running in Bandra, has revealed that the six accused who worked as waiters, managed to steal credit card and debit card data of as many as 1,000 customers from 96 different banks including eight major banks.

However, the Bandra police clarified that all of the stolen data may not have been used for stealing money. The data was stolen by the six waiters working in bars and restaurants using credit card skimmers at Mumbai, Pune and Thane from their unsuspecting customers and patrons since early 2016.

Three of them had worked at a restaurant called Persian Durbar in Sakinaka, Andheri. The unsuspecting customers would hand over their card as well as pin code to the waiters, who secretly swiped them on the skimmers. In some cases, they observed the customer while entering the pin code on the keypad and memorised it.

The Bandra police said that there are more links to the case and the involvement of more accused besides the 6 waiters has not been ruled out. Deputy commissioner of police, zone IX, Paramjit Dahiya told HT, We are in the process of identifying the people whose credit and debit card data was stolen.

Till now, 20 people have been identified and as many First Information Reports (FIRs) have been registered in Thane, Pune and Mumbai. The police are simultaneously also finding out if FIRs have already been registered by victims after realising that money had been withdrawn from their accounts.

Detection officer, sub inspector Bhimsen Gaikwad said, We had received a complaint from a private bank in May this year after which we traced the accused, which led to further arrests. Now, another major bank has approached us and we are taking their help to identify the victims.

The complaint filed in May by the private bank had said that an unidentified accused had withdrawn money from an ATM on Carter Road using a cloned card of a customer. The police worked out the information and arrested one of the accused this month. His arrest led to the arrest of seven others.

Apart from card skimmers, the police seized a laptop and a software they used to clone cards.The police have also seized 50 cloned cards which are re-writable. After using these cards to withdraw money, the accused wouls erase the used data and store new data on it by overwriting it.

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Card cloning racket: Mumbai gang steals debit, credit card data from 1000 customers of 96 banks - Hindustan Times

In the Future, You Will Have the Same Pet Your Entire Life – VICE

Near the end of my conversation with Jae Woong Wang, a researcher and spokesperson for South Korea's Sooam Biotech, he asks me to tell the world that they shouldn't stuff any recently deceased pets they hope to have cloned in freezers. It renders cell matter impossible to harvest, which isn't good news if you're in the business of cat and dog duplicates. It's hard to let a grieving family down easy, especially after they've made the day-long trip across the Pacific only to discover their newly dead companion won't be getting a genome-generated second chance.

"You have to preserve the body as long as possible without freezing," says Wang. "That's a mistake a lot of people make. When water freezes, it punctures all the cells, and the chances of cloning becomes extremely low. It's a frustration we're constantly dealing with."

Sooam Biotech's founder, Hwang Woo-suk, ran into significant controversy in 2004 when he fraudulently claimed to have cloned human embryos, but the company has stayed in the business for over ten years. Sooam has fulfilled contracts with the commercial farming industrycloning livestock for breeding and bottom-line purposesbut its pet cloning division is a marketplace built on a more spiritual communion. It'll take $100,000 to reunite with a reincarnated version of an animal you loved.

Its cloning process is more straightforward than you might think. A Sooam clerk will meet you at the Seoul airport and retrieve a fingernail-length biopsy of your dead pet's flesh. A donor dog or cat is selected from the company's kennel. Their eggs are flushed out, gutted of their genetic information, and fused with DNA harvested from the biopsy. If the process works, the retrofitted egg is inserted into a surrogate mother. "Until the point where they actually meet the dog, [the customer] is in a very happy disbelief," says Wang. "But once we deliver the dog, they usually burst into tears."

The jury is still out on what a clone actually is. It's a conundrum that's raged ever since Dolly, the famous duplicated sheep, was brought into the world in 1996. Genetically, they'll be a mirror image of the source animal, an asexually wrought son or daughter built in the flash of nuclear transfer. But will the clone share the same emotions or personality tics? That's difficult to say. Research on cloned cows and pigs has shown distinct differences in personalityand even looksfrom the animal of origin to its clone.

As such, New York Magazine's Science of Us blog called pet cloning "a laughable, extravagant waste of money," when news broke last year that media tycoon Barry Diller and fashion mogul Diane von Furstenberg had their Jack Russell terrier cloned, even though the wealthy power couple seemed pleased with the two puppies they got as a result. And, in an interview with Scientific American, stem cell biologist Robin Lovell-Badge maintained that cloning a pet was, flatly, "stupid." "You're never gonna get Tibble back, or whatever," he added.

But companies like Sooam deal in loveor more specifically, the faint chance that you might love again. Because losing a dog or a cat is really goddamn rough. "A beloved pet is much like a family member," reads the pitch on ViaGen Pets, a Texas-based commercial cloning outlet that offers a pet-cloning service. "The unique life-enriching bond, the love and companionshipa truly special pet provides us a unique sense of comfort and life-enriching fulfillment that is nearly impossible to extend beyond your pet's natural lifespan. Until now."

It was a convincing enough argument for Doug and Michelle Shields, and their fluffy white Maltese, Guinevere. Gwen lived 16 and a half years before she died after a seizure. The Shields had mulled the idea of preserving her genes in the past, but it wasn't until the fresh aftermath of her death that they made the decision to start the cloning process. (Luckily, the veterinarian put Gwen's carcass in a refrigerator, not a freezer or a cremator.)

"We're what you'd deem to be animal people. We have a parrot and another dog we adopted," says Michelle. "But Gwen was just an amazing, amazing, amazing dog. Just unbelievable. She just had a personality. Everyone loved her. There was no replacing her. So if I could get her back, or her personality traits, I would do anything to do that."

The Shields reached out to PerPETuate, an animal genome preservation business run by Ron Gillespie, who used to work at the cattle genetics company ABS Global. Right now, he's partnering with ViaGen, and recently, its laboratory delivered four clones sourced from a genome Gillespie first harvested in 2000. He happily preserved Gwen's DNA, and the Shields family is currently deep in the cloning process, one Gillespie remains optimistic about.

"[Customers] see the whole procedure as a healing journey."

"Dog owners [throughout history] have said, 'This is the best dog I've ever had,' and I'm going to breed them with another dog to get a puppy that's as closed to [the original] as possible," Gillespie says. "That's a very natural, common thing. Selective breeding has been going on for years. This is the ultimate breeding tool. You're not just getting half of the genes; you're getting 100 percent of them. It's an understandable step in the evolution of breeding."

Gillespie's currently working with a client with an autistic son who finds peace in the presence of an old family cat. The client is, of course, terrified of what might happen after the cat dies. He's tried other animals (and other cats), but nothing musters the same pacifying effect. So instead, he holds out hope that maybe he can give his son some peace with a long line of duplicates.

"This cat is of exceptional value to this boy and to this family," says Gillespie. "They tried the brother of this cat, and the boy totally rejected his brother. So they're going to clone him. And what's gonna happen? Is this cat gonna be able to substitute? If it is, think about the significance of that. People don't think in those terms. They just think it's just rich people with a lot of money."

Gillespie tells me that 2017 has been one of the busiest years for PerPETuate since he started the business in '98, and speculates eventually pet cloning will become more common as the prices get more affordable. He thinks ViaGen is a good first step, as they offer a cloning service domestically for $50,000a bargain compared to Sooam's six-figure entry fee. But obviously, that price will have to come down quite a bit more for cloning to truly hit the mainstream.

In a way, Gillespie has been banking on cloning become more accessible for the entirety of his career, since his business is basically built around preserving genomes for an era where it does become more economically viable for the average pet lover. Until then, it's not just the Diane von Furstenbergs of the world who are writing big checks to bring back their beloved pooches: As of the fall of '15, Sooam Biotech estimated it had cloned some 600 dogs, not all of which had wealthy owners. Jae Woo Wang tells me some of their customers liquidate assets to afford the cloning process. Priorities tend to shift in the midst of grieving.

ViaGen's testimonial section illustrates that, no matter the cost, its customers believe it's money well spentdozens of former clients there have drafted sonnets in tribute of the preserved genomes of their dead pets and the hopes for the possible clones they may one day produce. "They see the whole procedure as a healing journey," says Gillespie. "At first it's very difficultyour dog just passed away, you have to go to a vet to get a biopsy done and send it over, or sometimes travel here. You have to wait for us to give the confirmation that the cells are OK. All of that is very, very stressful. But once they actually have the puppy, that's when they unload."

Michelle and Doug Shields just want their dog back, and look at the price tag as a worthy luxury. Some more time with Gwen is a far more important splurge to them than a trip to Italy or a Country Club membership. Michelle says that most of her friends understand, because they all loved Gwen too. "We're just people who really love our dog," Michelle says.

Follow Luke Winkie on Twitter.

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In the Future, You Will Have the Same Pet Your Entire Life - VICE

This powerful new tool for cloning genes could speed up drug discovery – Boston Business Journal


Boston Business Journal
This powerful new tool for cloning genes could speed up drug discovery
Boston Business Journal
The technology "makes a 20,000-piece puzzle look like a 1,000-piece puzzle, said one of the researchers, Biju Parekkadan, a faculty member at Massachusetts General Hospital. Subscribe to get the full story. Already a subscriber? Sign in. Subscribe to ...

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This powerful new tool for cloning genes could speed up drug discovery - Boston Business Journal

Yves Tumor’s New Music Video is a Creepy Story of Cloning & Biohacking – Out Magazine

Queer musician Yves Tumor's new film, for the frenetic track "Broke In," is a chilling exploration of cloning, biohacking, and being buried alive. For the project, Tumor tapped Berlin filmmakers Sam Dye and Bliss Resting to direct, with lyrics by UK musician Oxhy.

Its about feeling crushed by the city of London, walking around the streets and feeling no escape, feeling like the city youre from wants you out, Oxhy told Dazed. Needing release, but at the same time constantly remembering the violence that the desire for freedom as a male has meant, historically and currently.

Much of the film sees Oxhy covered in dirt, with just his eyes and his muddy teeth visible. The result is a highly jarring, unsettling film that includes images of great violence and oppression.

Oxhy is the real star of this film, said Dye and Resting.We put him through so much, its fantastic to have such dedicated people to work with. Always a star, no matter how much hes choking.

Tumor's debut album,Serpent Music, came out last year, and impressed us with its experimental, atmospheric approach to creating a queer soundscape. Tumor is based in Turin, Italy, and an affiliate of another highly important LGBTQ musician, Mykki Blanco.

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Yves Tumor's New Music Video is a Creepy Story of Cloning & Biohacking - Out Magazine

Three waiters arrested in Mumbai for fraud and cloning ATM cards … – Hindustan Times

The Bandra police recently arrested three more waiters working for a hotel in Sakinaka for allegedly using skimmers to steal credit and debit card details of their customers to cheat them later. With this, the total number of arrested accused in the case reached eight.

DCP, zone IX, Paramjit Dahiya said, Eight people, including six waiters, have been arrested. Eight skimmers and 50 ATM cloned cards have been seized.

Apart from Bandra police in Mumbai, FIRs have been registered in Thane and Pune districts for duping unsuspecting customers. According to Bandra police, the three waiters were working in Persian Durbar in Sakinaka area in Andheri east. Two other waiters one from Pune and another from Thane were also arrested. The unsuspecting customers coming over to the Sakinaka hotel not only handed over their credit card/debit card to the three waiters but also gave their PIN code to them.

The waiters swiped the card on the three skimmers that have been recovered from them. The stolen information was then used to clone cards with the help of a software and laptop, said detection officer, sub inspector Bhimsen Gaikwad from Bandra police station. Santosh Shetty, general secretary of Association of Hotels and Restaurants (AHAR) having 8,000 hotel establishments as members in Mumbai told HT, We have put up sign boards at our hotels as well as we ask our waiters to go up to the table with the payment machine. We ask customers to swipe the card and enter the pin code with their own hands. In general, customers should desist from handing over their card and pin while making payment at anyplace.

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Three waiters arrested in Mumbai for fraud and cloning ATM cards ... - Hindustan Times

Puppies cloned from ears arrive in Russia for genetic research … – RT

Published time: 24 Jun, 2017 08:49 Edited time: 24 Jun, 2017 10:16

Lab-produced dogs cloned from bio material taken from the two best representatives of the Yakutian Laika species have arrived in Russia from South Korea for genetic research. One met its original mother as a Ruptly crew filmed the moment.

The puppies, which were cloned by South Koreas Sooam Biotech Research Foundation, one of the worlds leading dog cloning laboratories, arrived in Yakutsk, the capital of Siberias Yakutia Republic, on Friday. The research was led by Dr. Hwang Woo Suk and his team.

The scientists succeeded in cloning the Yakutian Laikas, a type of hunting dog from Northern Russia and Russian Siberia, from a 12-year-old male and a 6-year-old female.

One of cloned puppies is a two-month-old girl named Kyrachana, which means beautiful in the Yakutian language. Another is a 3-month-old boy named Belekh, which means present.

Ruptly visited the farm where Kyrachana is now living when she met her original mother for the first time.

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I cant really believe that it [the puppy] is a clone. If you look at the original, you can see that they both look alike,the dogs owner Dmitry said, adding that the puppy had been cloned from a part of the mother dogs ear.

The collaboration with the South Korean scientific laboratory is aimed at saving the original Yukutian Laika breed, whose population has seen drastic decline over past decades due to cross-breeding.

The canines were cloned in the Sooam Biotech Research Foundation, which is focused on advanced biotechnology for industrial and biomedical applications using animal cloning and pluripotent stem cells combined with transgenic technology, the companys website says.

However, not all of the cloned dogs are pets. The laboratory has replicated the best military and police dogs to assist the South Korean police, and many cloned dogs currently work in police forces in the US and China.

In November of 2016, the company sent three cloned Belgian Malinois to join police forces in Yakutsk.

Animal cloning is not allowed everywhere in the world. In 2015, the EU Parliament banned the cloning of animals, as well as importing their descendants.

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Puppies cloned from ears arrive in Russia for genetic research ... - RT

Magnified: Cloning – The Hawk Eye (blog)

Cloning: gathering a persons DNA to make a replica of them. This topic can go into more depths than just the first cloning case of Dolly the sheep. Cloning is often related to the Illuminati and people they are trying to reach. Many conspiracy theorists believe clones of celebrities are used to influence the public in a mind controlling way.

Physical attributions and actions of many celebrities have changed over the years. These changes of famous celebrities are believed to show cloning in action. Celebrities are known to have experienced many cloning glitches during their performances or daily lives.

Does the Illuminati really have anything to do with all this cloning drama? Many theorists, including myself, believe that many celebrities have been killed by the Illuminati and are being replaced by clones. The ever rising list of these celebrities include Eminem, Al Roker, Beyonc and more.

The Illuminati sends out small symbols which are connected to the organization like triangles, fake satanic-like rituals, the eye, and snakes. You can see this through many music performances of artists like Ariana Grande, Beyonc, Katy Perry, Michael Jackson, and many more. These signs can also be found in necklaces, rings, cloaks, and any attire. Many celebrities have been seen showing the ever famous triangle with their hands.

By killing, cloning, and replacing these celebrities, the Illuminati is subtly influencing the public. They are changing the government in order to promote the New World Order. This group wants to infiltrate other organizations and make people act like their slaves.

Beyoncs actions during a basketball game show us a possible zoned-out clone, almost looking as if she was receiving instructions. There were alleged claims that she was killed and then cloned to prevent the possible mass hysteria her death could bring.

The same glitch occurred to news reporter Al Roker during a news story. His fellow reporters said the words Holy Ghost and he started staring into the camera. This very creepy look on his face stayed for a good 50 seconds, which is not a normal thing to occur during a broadcast story. This was later reported as just a bet between the producer and Roker, but they could just be covering up the possibility of cloning.

This glitch may have also affected the famous rapper Eminem. After being checked into rehab, he was believed to have died there and been cloned. This brought a lot of controversy to the topic and the legitimacy of Eminem.

All of these cloning incidents bring some humor into the whole conspiracy world. I mean, come on, look at those faces.

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Magnified: Cloning - The Hawk Eye (blog)

How Close Are We to Successfully Cloning the First Human? – Futurism

When Will We Clone a Human?

Human cloning may endure as one of the go-to science fiction tropes, but in reality we may be much closer to achieving it than our fictional heroes might imply. At least in terms of the science required. On of the most prominent hurdles facing us may have less to do with the process and more to do with its potential consequences, and our collective struggle to reconcile the ethics involved.That being said, while science has come a long way in the last century when it comes to cloning a menagerie of animals, cloning humans and other primates has actually proven to be incredibly difficult. While we might not be on the brink ofcloning entire human beings, were already capable of cloning human cells the question is,should we be?Click to View Full Infographic

The astoundingly complex concept of cloning boils down to a fairly simple (in theory, at least) practice:you need two cells from the same animal one of which is an egg cell from which youve removed the DNA. You take the DNA from the othersomatic cell and put it inside the devoid-of-DNA egg cell. Whatever that egg cell goes on to produce for offspring will be genetically identical to the parent cell.While human reproduction is the result of the joining of two cells (one from each parent, each with their own DNA) the cellular photocopy technique does occur in nature.Bacteria reproduce through binary fission: each time it divides, its DNA is divided too so that each new bacterium is genetically identical to its predecessor. Except sometimes mutations occur in this process and in fact, that can be by design and function as a survival mechanism. Such mutations allow bacteria to, for example, become resistant to antibiotics bent on destroying them. On the other hand,some mutations are fatal to an organism or preclude them coming into existence at all. And while it might seem like the picking-and-choosing thats inherent to cloning could sidestep these potential genetic hiccups, scientists have found thats not necessarily the case.

Image Credit: Pixabay

While Dolly the sheep might be the most famous mammal science has ever cloned, shes by no means the only one: scientists have cloned mice, cats, and several types of livestock in addition to sheep. The cloning of cows has, in recent years, provided a great deal of knowledge to scientists about why the processdoesnt work: everything from implantation failure to those aforementioned mutations that render offspring unable to survive.Harris Lewin, professor in the UC Davis Department of Evolution and Ecology, and his team published their findings on the impact cloning has ongene expression in the journalProceedings of the National Academy of Sciencesback in 2016. In the studys press release Lewin noted that the findings were certainly invaluable to refining cloning techniques in mammals, but that their discoveries also reinforce the need for a strict ban on human cloning for any purposes.

The creation ofentiremammals via reproductive cloning has proven a difficult process both practically and ethnically, as legal scholar and ethicist Hank Greely of Stanford University explained toBusiness Insiderin 2016:

The cloning of human cells,however, may be a far more immediate application for humans.Researchers call it therapeutic cloning, and differentiate it from traditional cloning that has reproductive intent. In 2014, researchers created human stem cells through the same cloning technique that generated Dolly the sheep. Because stem cells can differentiate to become any kind of cell in the body, they could be utilized for a wide variety of purposes when it comes to treating diseases particularly genetic diseases, or diseases where a patient would require a transplant from an often elusive perfect match donor.This potential application is already well underway: earlier this year a woman in Japansuffering from age-related macular degeneration was treated with induced pluripotent stem (iPS) cells created from her own skin cells, which were then implanted into her retinas andstopped her vision from degenerating further.

We asked the Futurism community to predict when they think well be able to successfully clone a full human, and the majority of those who responded agree that it feels like were getting close: nearly 30 percent predicted well clone our first human by the 2020s. We have replaced, and replicated almost every biology on earth, said reader Alicja Laskowska, [the] next step is for cures and to do that you need clean DNA, and theres your start.

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How Close Are We to Successfully Cloning the First Human? - Futurism

Cloning To Revive Abaco Wild Horses – Bahamas Tribune

Some of the wild horses in Abaco.

ON April 4, the government approved a proposal to restore life to an extinct sub-breed of Spanish horses through cloning, according to a press release from the Wild Horses of Abaco Preservation Society (WHOA).

The WHOA said government approval of the proposal to initiate the cloning process and to bring the horses back to Abaco enables it and another group, Arkwild Inc -- a charity in the US -- to begin major fundraising to make the restoration a reality by restoring the horses' preserve to excellent condition prior to the arrival of the clones.

"The Abaco island horses thrived on the island of Abaco...when they were imported in the late 1800s," the press release noted. "Human intrusions and an increasingly toxic environment led to a decline. A 25-year effort to save the horses ended on July 23, 2015 when the last mare died.

"Living tissue was saved from that last mare and in a dramatic series of events --worthy of an international suspense thriller -- the tissue was airlifted to the US where ViaGen, a Texas firm engaged in cloning, found the cells viable and nurtured enough to clone two mares.

"Pro bono services valued at close to $2m have been pledged to the cloning effort by companies and individuals in the US."

The group also explained its plans for the horses' preserve.

"Along with the dramatic restoration of the herd, the horses' preserve will become a showcase for equine assisted therapy, alternative energy systems, ecotourism and many related activities consistent with UN (United Nations) sustainable development goals.

"In an environment that is unique in the world, a unique horse once again will roam the pine forests of Abaco, 200 miles out at sea.

"Genetically unique after over a hundred years of isolation on Abaco, the Abaco horses were direct descendants of the distinctive horses of the Conquistadores who led the first major forays into the 'New World.' Those June 12, 2017 horses could best be described as 'Jeeps,' being tough, steady, sturdy and needing far less water and feed than standard domestic horses," the preservation society noted.

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Cloning To Revive Abaco Wild Horses - Bahamas Tribune

Cloning to the rescue – New Scientist

Having cloned sheep and who knows what else, wouldnt it be a good idea for scientists to begin cloning the worlds endangered species? Is there any reason not to?

Cloning technology has improved dramatically since 1996 when Dolly the sheep became the first mammal to originate from a somatic cell (any cell of a living organism other than the reproductive cells). We now have a better understanding of nuclear reprogramming, a process that gives a cell the capacity to generate all the different types of cells that make up the organism called totipotency. As a result, many species have been successfully cloned, including those of amphibians, fish, insects and mammals. But cloning is still inefficient, with a maximum of 5 per cent of cloned embryos developing into healthy offspring.

There have been several attempts at cloning endangered or even extinct species, such as the gaur and the Pyrenean ibex. These have been largely unsuccessful. Generally, hundreds of embryos have to be created, of which only a handful can be implanted. And of those, only a couple are born, but these often die soon after birth.

The problem in most cases is sexual isolation, which is part of the speciation process, in which one species eventually evolves into two separate ones. These become incapable of reproducing with each other because their reproductive cells are no longer compatible or the embryo of one species cannot be carried by a mother of the other. Both issues affect cloning.

One species where cloning has been successful is the African wildcat. Although wild, this species is still genetically close enough to the domestic tabby for interspecies embryo transfer to be slightly more efficient. More importantly, cloned animals were then able to mate and reproduce among themselves.

But the main problem with cloning remains: it eliminates genetic diversity. The whole point is to reproduce an exact copy of an individual, such as a champion racehorse. Most higher organisms, however, reproduce sexually, which leads to the offspring bearing a combination of characteristics from both parents. This genetic mix and match allows species to adapt to their environment and reduces negative traits or diseases within the population. A diverse genetic pool is essential for a species to survive in the wild.

So although cloning might help preserve some species in critical times, the best way to avoid their extinction is to protect their environment and stop senseless poaching of wild animals.

Alena Pance, Wellcome Trust Sanger Institute, Cambridge, UK

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Cloning to the rescue - New Scientist

Hackers caught cloning activist Twitter accounts to spread fake news – The Independent

Designed by Pierpaolo Lazzarini from Italian company Jet Capsule. The I.F.O. is fuelled by eight electric engines, which is able to push the flying object to an estimated top speed of about 120mph.

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A humanoid robot gestures during a demo at a stall in the Indian Machine Tools Expo, IMTEX/Tooltech 2017 held in Bangalore

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A humanoid robot gestures during a demo at a stall in the Indian Machine Tools Expo, IMTEX/Tooltech 2017 held in Bangalore

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Engineers test a four-metre-tall humanoid manned robot dubbed Method-2 in a lab of the Hankook Mirae Technology in Gunpo, south of Seoul, South Korea

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Engineers test a four-metre-tall humanoid manned robot dubbed Method-2 in a lab of the Hankook Mirae Technology in Gunpo, south of Seoul, South Korea

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The giant human-like robot bears a striking resemblance to the military robots starring in the movie 'Avatar' and is claimed as a world first by its creators from a South Korean robotic company

Jung Yeon-Je/AFP/Getty Images

Engineers test a four-metre-tall humanoid manned robot dubbed Method-2 in a lab of the Hankook Mirae Technology in Gunpo, south of Seoul, South Korea

Jung Yeon-Je/AFP/Getty Images

Waseda University's saxophonist robot WAS-5, developed by professor Atsuo Takanishi

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A test line of a new energy suspension railway, resembling a giant panda, is seen in Chengdu, Sichuan Province, China

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Japan's On-Art Corp's eight metre tall dinosaur-shaped mechanical suit robot 'TRX03'

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Japan's On-Art Corp's eight metre tall dinosaur-shaped mechanical suit robot 'TRX03' performs during its unveiling in Tokyo, Japan

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The interior of Singulato Motors' concept car Tigercar P0 at a workshop in Beijing, China

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Singulato Motors' concept car Tigercar P0

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A picture shows Singulato Motors' concept car Tigercar P0 at a workshop in Beijing, China

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An exhibitor demonstrates the AnyWalker, an ultra-mobile chasis robot which is able to move in any kind of environment during Singapore International Robo Expo

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Government and industry are working together on a robot-like autopilot system that could eliminate the need for a second human pilot in the cockpit

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Hackers caught cloning activist Twitter accounts to spread fake news - The Independent

Cloning Grapes Will Save Australian Wine – National Geographic Australia

Australian grapes are unique. So unique they have the potential to surpass other new world wines such as popular varieties found in America.

Michael McCarthy, Primary Research Scientist for Viticulture at South Australias Research Development Institute, has been testing cloned grapes in Australias warmer wine territories. The aim of the experiment is to test how the cloned grapes will react to warmer climates particularly when temperatures in Australia are expected to rise dramatically in 50 years time (due to climate change). McCarthy explains the importance of the experiment:

Maybe the rest of the world might be more interested in some of our material. We have clones that just don't exist in the rest of the world anymore because our planting is clean. Phylloxera is not an issue, root-borne virus transmission is not an issue. We have planting material in Australia that is probably unique to the rest of the world.

McCarthy discussed the issue with vignerons operating in colder climates in Orange and central west New South Wales. The grapes cloned in one region may work just as well in other regions, saving many of Australias cooler wine regions.

[We are] understanding how wine styles within regions may change as that region warms up going into the future. What we are trying to do is identify sites across Australia that have the same clones in common. We take out the clonal difference and look at the wine style from cool to hot regions. So if this currently cool region becomes a warm region in 50 years' time how will those wine styles change?

The area in question is famous for Chardonnay which happens to be Australias number one white wine export, despite a decline of Australian consumption.

The call for Australian wine overseas, particularly our Chardonnay, is evident. As recently as last year 765 million litres of Australian wine was sent abroad valuing at $2.2 billion. According to trade minister Steve Ciobo:

Unlike other countries, Australias winemakers are not restricted by rules governing what they can plant or the types of wines they can produce. This has allowed innovation to thrive, driving Australias reputation for quality, consistency and diversity

China was revealed to be Australian winemakers best customer, with the US, UK and Canada following.

The experiment will hopefully enable winemakers to continue growing quality grapes despite the predicted rise in temperatures in the next 50 years.

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Cloning Grapes Will Save Australian Wine - National Geographic Australia

Five Rules For Successful Marijuana Cloning

Marijuana growers have two choices when it comes to starting new plants: grow from seed, or grow from clone.

Nowadays, its easier to buy marijuana clones than ever before, especially if youre in Colorado or Washington.

And marijuana clones do offer advantages that growing cannabis from seed cant offer

Growing from clones youre sure to have allfemales, and as long as you do the cloning yourself, you know who the mother plant is and how your clones will grow, yield, and get you high.

But theres an art and science to marijuana cloning, and weve devoted several articles to it. In this one, we give you five rules for more-successful marijuana cloning:

Take your cannabis cuttings from a healthy motherplant.Cloning gives yougenetic duplicates of your marijuaan motherplant.

If your motherplants are strong, healthy, and have potent genetics, your clones are getting the best start possible.

Avoid using motherplants that are sick, or that are plagued with pests. The only exception to this is if the sickness or pests can be purged from the clones using interventions.

In general though, if the mothers are bad, the clones will be too.

Give Your Marijuana Clones a Consistent Climate.Most cannabis clones prefer temperatures slightly higher than your ideal grow room temperature, and they dont like a day and night cycle with a temperature drop at night.

Thats why most growers root their cuttings under fluorescent lighting that stays on 24 hours a day. The lighting itself creates temperature stabilization.

Temperatures between 74-77Fahrenheit are in the range you want.

If you go colder than that, the cuttings root development might stall or just never happen at all, especially if your cutting root zone is below 69 degrees F.

You can increase your marijuana cuttings root zone temperature by using a propagation heating mat.

If youre growing in true hydroponics like deep water culture or aeroponics, use an aquarium heater to warm up your water temperatures.

If your cuttings root zone goes much warmer than 79 degrees F though, you run the risk of creating an environment favorable to damping off or root rot microorganisms that can ruin your cuttings.

In that case, you get a chiller, or you control clone environment so temperatures in the root zone are in the ideal range.

Clones like high humidity: Your cuttings need a humid environment in the 75-95% humidity range until theyve created roots and are able to intake water through them.

Thats why many marijuana cloners use humidity domes. Other growers say the domes can create root rot or stem root conditions, so they prefer to mist their cannabis clones rather than putting a dome over them.

The best domes for marijuana cloning have aeration vents built in. You want to un-dome or ventilate your cuttings several times a day so they get fresh air.

Just remember, with clones its best to go for too much humidity rather than not enough. Until your clones have functioning roots, if theyre in dry air, theyll die.

Use kind hydroponics lighting. You can use high-output fluorescent, LED, or plasma lighting for optimal marijuana cloning. HID hydroponics lighting is too intense for clones.

Keep your lights on 24 hours a day, and make sure that your clones arent too close to the bulbs or LED chips. Clones can easily burn.

Use sterile equipment and materials.You already figured out that your scissors, razor blade, or other tool for cutting clones has to be clean.

But you also want to make sure you trays, cloning domes, and root zone media are clean and sterile too.

Dont re-use cloning cubes or cloning powders and gels. Start fresh every time. You can read more about cloning powders and gels in this really useful article.

When you follow these marijuana cloning tips along with the tips in the other cloning articles linked within this article, your clone success rate should be nearly 100%.

If youre taking 20 clones and seeing only 10 of them survive, somethings wrong, and you want to closely re-evaluate your cannabis cloning techniques, the equipment and environment you use for cloning,and your marijuana motherplants.

We want you to enjoy 100% marijuana cloning success!

Aeroponics, Cannabis, Cloning, featured, Fluorescent, Growing Medical Marijuana, HID, Humidity, LED, Lighting, marijuana seeds, Mother Plant, Rooting, Temperature

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Five Rules For Successful Marijuana Cloning

Skimming, cloning become popular in Tulsa – KRMG

In prepared testimony for the Senate Intelligence Committee, James Comey will detail a series of meetings and phone conversations with President Donald Trump in 2017, as the former FBI Director says that he felt pressured by Mr. Trump to end an investigation of top Trump aide Michael Flynn, and that the President repeatedly asked the FBI to tell the public that he was not under investigation. You can read the full testimony from Comey, which was released by the Senate Intelligence Committee. Here are some of the details from the former FBI Director: 1. Comey: President Trump asked him for loyalty. At a January 27 dinner that involved only the two of them, Comey said he told Mr. Trump that as the FBI Director, he was not on anybodys side politically. Comey quotes the President as saying soon after, I need loyalty, I expect loyalty. That was followed by an awkward silence, according to Comey. 2. Comey: Trump asked him to drop Flynn investigation. The former FBI Director says that after attending a February 14 Oval Office meeting with other top officials, he was asked to stay behind by the President, who quickly made clear the topic. I want to talk about Mike Flynn, Comey quotes Mr. Trump, in talking about the investigation of Flynn, who had just resigned as the Presidents National Security Adviser. I hope you can see your way clear to letting this go, to letting Flynn go. He is a good guy. I hope you can let this go, Comey quotes Mr. Trump. 3. The cloud of the Russia investigation. The next conversation recounted in Comeys testimony occurred on March 30 in a phone call, as Comey says the President referred to the Russia probe as a cloud over his presidency. During that meeting, Comey says Mr. Trump asked multiple times for the FBI to publicly say that there was no direct investigation of the President. He repeatedly told me, We need to get that fact out,' Comey recounted. 4. More concern about the Russia cloud. The last conversation between the two men was also by telephone on April 11. Comey says the President asked why there had not been any announcement that he was not under investigation, as Comey said he was told that the cloud was hampering his work as President. 5. There were other conversations not detailed. In his testimony, the former FBI Director says he can recall nine one-on-one conversations with President Trump in four months there in person and six on the phone. This testimony does not go through all of those there is no indication given as to why those were not included. BREAKING: Comey to tell Senate committee he found Trump request to end Flynn investigation 'very concerning.' AP Politics (@AP_Politics) June 7, 2017

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Skimming, cloning become popular in Tulsa - KRMG