The process in nature or in the lab by which a new organism is created that is genetically identical to its predecessor.

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Does a woman contain all the genes needed to make a man?

I know the answer of inverse is at here, but how about this question? I also read this question, can I imply to human that a woman can also contain all the genes needed to make a man? Edit: I ...
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1answer
21 views

What is the current status of human cloning?

Human cloning is a very controversial subject. I had a professor at Uni whose subject was transgenics. He had a very successful career both in industry and in academia. We were kinda close and he ...
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11 views

Requirements for In-Fusion Cloning Protocol?

I have some long primers I'd like to repurpose for an In-Fusion cloning project. The In-Fusion manual can be found here. They list a few examples of how the primers should match the vector, with 15 ...
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37 views

Sequence Scan for CRISPR- sgRNA design & scoring question [on hold]

My lab is attempting to implement the Sequence Scan method from the publications "Sequence determinants of improved CRISPR sgRNA design" We have used the website referenced in the publication, ...
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1answer
22 views

Methods for varying insert size in E. coli clones

I'm writing my master's thesis proposal, and I need to improve the solution-behavior of some truncated protein domains which have previously shown poor solubility when expressed in E. coli. To ...
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1answer
21 views

how do we prevent restriction enzyme of bacteria to cut and destroy our new recombinant?

Based on cloning principle, the new DNA recombinant will be introduced to host cells, commonly bacteria, how do we prevent restriction enzyme of bacteria to cut and destroy our new recombinant ?
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67 views

Are identical twins clones? [closed]

Identical twins have the same genetic make up but they differ in development so are they in a way clones?
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26 views

What's the rationale behind freezing one's eggs if cells can be cloned?

It seems to me there's a lot of discussions around the topic of somebody freezing her eggs. What I don't understand though are the following: Why would you freeze your eggs when you know that the ...
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25 views

UV light-induced mutagenesis during gel extraction

this is a very short question I did not find the answer for online, neither on this nor other fora. At the beginning of my cloning protocol, I extracted the band with the sequence of interest directly ...
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1answer
22 views

Using PGAL10 promoter to overexpress specific yeast genes: How to?

I was reading a poster about expression boosting for Artemesinin production in yeast and it had this line: PGAL1 and PGAL10 have been used for the overexpression of all genes from the ...
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17 views

de novo gene synthesis vs cDNA library creation from plant RNA: Pros & Cons

What are the relative pros & cons of the following two approaches when one wants to insert a known, sequenced plant gene into a target host organism: de novo gene synthesis vs plant matter ...
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1answer
50 views

What are the problems in using an ssDNA or an mRNA for expression instead of using a plasmid?

Let's say you want to do a gene cloning experiment. You have an empty expression vector (mRNA or DNA) and are looking to clone some gene X into it. Why is it you can only buy X already inside a vector ...
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1answer
21 views

Unwanted E. coli product in plasmid product

Using a TOPO XL Cloning kit, our lab took some amplified human DNA, inserted it into the vector, and attempted to clone it in E. coli. However, upon sequencing the purified plasmid product, the ...
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1answer
43 views

Cloning Ice Age Lion Cubs

It was just on the news that scientists discovered "Two cubs in Russia's Sakha Republic last August in a near-perfect state". And the South Koreans and Russians are attempting to clone these extinct ...
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2answers
250 views

How does LCR compare to Assembly PCR

The question pretty much explains itself. How do the two methods compare? I've always used Assembly PCR but the method is prone to mistakes and I'm curious how it compares to Ligase Chain Reaction ...
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0answers
18 views

codon optimization / enzyme active site improvement

Does one necessarily have to use de novo synthesis of DNA when attempting protein expression improvement by codon optimization? Or are there other ways? e.g. Say the gene coding for the enzyme needed ...
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16 views

Cloning Stragedy

Recently I am learning about TOPO cloning in a class. I have questions regarding to this: In the TOPO-TA cloning, they use topoisomerase I to cut the backbone of the vector so it will covalently bind ...
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54 views

What tools are used in animal cloning?

So I was reading up on articles to do a project about cloning but there were no places in the article where It states the tool used to take the gene out of the nucleus and insert it into plasmids ? ...
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2answers
169 views

In Sanger sequencing, why do we resort to cloning? Why doesn't PCR suffice?

I understand that in Sanger sequencing we can clone our fragments with the help of e.g. bacteria to make multiple copies of our fragments for further analysis. I also understand cloning can be a ...
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3answers
2k views

Does a man contain all the genes needed to make a woman?

This question is brought on by a Sci Fi novel I am thinking about writing. The plot device involves a colonist in charge of building a population on a new planet who loses his supply of embryos and so ...
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2answers
57 views

Alternatives to His-tags

What are the alternatives to using a histidine-tag in detecting protein-protein interactions? Why would they be more beneficial than his-tags?
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37 views

What is the sequence of pPROLar.A plasmid?

I've been looking for the sequence of pPROLar.A122 plasmid but I haven't found it. Where could I find the sequence? The first link (addgene) on googling reports a plasmid which is reported to have ...
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0answers
15 views

What will happen if an anaplastic/cancer cell is used in cloning?

Since anaplastic cells have high glycolytic ratio and replicative potential, if they are used as the somatic cell in a model of cloning(like dolly's mother's udder cell), what is likely to happen?
4
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1answer
146 views

Freezing after restriction digest

Brief question: I have performed a double digest with heat inactivated restriction enzymes with no star activity. I usually purify the DNA (PCR inserts and linearised vector) using a kit or gel ...
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1answer
58 views

Are progenies factually half-clones of the parents?

Given that a "clone" (if anything, in the context of human cloning) is taken to be, as far as I have understood, a specimen possessing the same genome as his "father/mother", aren't all "non-cloned" ...
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30 views

“Antibiotic resistance” equivalent in archaea for selection during cloning

I'm beginning to work with halophilic archaea and I'm trying to figure out a good way to select for cells that have taken up a plasmid. Obviously, one can't use antibiotic resistance since they are ...
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36 views

Plasmid with ITPG and pBR322 ori

I'm looking for suggestions as my internet searches aren't turning up much. I'm trying to find a plasmid that has the pBR322 origin, as it's low(ish) copy number and can tolerate large inserts. ...
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2answers
273 views

How can we create a living dinosaur using DNA technology?

I'm wondering what would we need to do to create a living dinosaur using DNA technology? If it is not possible with current technology, will it ever be feasible? In the movie Jurassic Park, ...
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1answer
277 views

Why are female clones more often produced

As a student of biology when ever I come by artificial cloning, I always find examples of females being cloned - Dolly the sheep, CopyCat, Daisy, etc. The only male I could see was Fibro mouse and a ...
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1answer
304 views

How bad is ethidium bromide in your plasmid for downstream applications?

I want to transfect Adipose Derived stem cells using a clone of my own. My problem is that I get a lot of contamination (not only genomic, but due to the nature of the technique, I get undesired ...
3
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2answers
262 views

Neomycin resistance gene in E. coli stb3 cells

We use a lentivirus packaging vector called pLenti_puro_DEST. We use puromycin to select for the cloned gene in 293T cells. However, the pLenti_Neo_Dest has been discontinued, so I'm cloning a ...
3
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1answer
225 views

Is the amount of dNTPs rate limiting for very long PCR products?

I'm using the Q5 system and I'm PCRing a product that will have a final length of around 9500 bases. I have noticed that the product is there but very faint. The primer seems to be used up. So my ...
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75 views

Closing a Plasmid by Ligation

I have been having trouble with what should be a fairly simple ligation. I had a plasmid that I needed to cut part out of, so I designed a couple PCR primers to do that. The plasmid already had a ...
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1answer
141 views

Does DNA Ligase look for complementarity in sticky ends?

Can DNA ligase seal two non-complementary sticky ends if the reaction is incubated at the optimum conditions ? I do almost 100% double digestion in the lab. However, during ligation, when I do a ...
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59 views

How Many Generation We can Run a Mycelium of Pleurotus ostreatus/Pearl Oyster?

I am able to grow mycelium successfully on the spent coffee grounds. And mycelium is successfully fruiting once/twice on the same substrate. I am cloning by taking mycelium from the existing one ...
3
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1answer
709 views

Open protocol for Ligase Independent Cloning

Ligase Independent Cloning is a protocol that allow an insert to be integrated into a vector without ligation. It uses T4 DNA polymerase with only ATP to first chew back from blunt ends to create ...
3
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1answer
61 views

Literature about putative epigenetic state changes in mammal sequences after cloning steps in Escherichia coli

I would like you to point me out some literature about putative epigenetic state changes in mouse/mammal sequences after cloning steps in Escherichia coli. This are the last search details I used in ...
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1answer
150 views

Molecular Cloning- Blunt end restriction endonucleases

I work in a microbiology lab where we do a lot of cloning. I have always used restriction endonucleases to cleave the DNA to have sticky ends and not blunt ends. I currently am working on a project ...
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1answer
598 views

Cloning and Telomeres

I read that the length of telomeres in Dolly was shorter, because the DNA was taken from an old sheep. Shorter telomeres may lead to early aging. What is happening now with cloning? Do we use a ...
13
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1answer
193 views

Organ cloning - possible to make a non-antigenic organ?

From a J. Neil Schulman article on Organ Cloning: Cannibalizing organs from other people also entails the risk of rejection because of incompatibilities, not only for tissue-typing but also ...
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1answer
133 views

Possibility of Human cloning

Technology has evolved so much as to clone animals. But, what about human cloning? Is it really possible to create humans in lab? Or is it that technology has not advanced to that level..? Why cant ...
2
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1answer
65 views

Cloning a gene of an organism with an nonsequenced genome

What would be the best protocol to clone a gene about 5kb in size? The genome is not sequenced, but the gene itself very similar to orthologous genes of organisms with known sequences.
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2answers
107 views

Is there a command line molecular cloning tool?

There are many softwares such as SnapGene, Geneious, Vector NTI, ApE, Gentle and others for assistance with cloning. Their key functions are: Making maps of plasmids Annotating plasmids with common ...
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31 views

Gibson assembly - primer design with A and T rich regions

I have question about Gibson assembly. I have done it several times and it always worked okay for us, but now I want to assemble together a fragment which has sequence like this: ...
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0answers
50 views

What should be the distance between plant promoter and gene?

I am trying to clone a rice gene under a different endogenous rice promoter.I will be cloning the CDS of the rice gene.So I wanted to know what is the minimum or maximum distance I should put between ...
3
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1answer
894 views

How much DNA of Albert Einstein is recoverable?

Since there seem to be five biological descendants of Albert Einstein, and the original chromosomes of him distributed among them ... Is it possible to recover enough DNA of an individual from his ...
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923 views

Why doesn't T5 exonuclease have endonuclease activity in Gibson assembly?

Gibson assembly uses T5 exonuclease to chew back the 5' end of dsDNA to generate overhangs. However, T5 exonuclease, in contrast to lambda exonuclease, is reported to have ssDNA endonuclease activity ...
3
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1answer
9k views

What is a suicide plasmid?

What is a suicide plasmid? Example: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2737453/ mentions a "suicide vector". See also ...
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1answer
186 views

What are the Gateway clonase enzymes?

The Gateway cloning system utilizes what Life Technologies refers to as "Clonase enzyme mix" to catalyze the BP and LR reactions. What is in this enzyme mix? Is there a sequence for them?
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How to DIY preserve pet DNA today so that it can be used in 20 years

I know that there are companies that offer a kit+storage of your pet DNA for around $2k. My question is if there is any other option for doing this yourself with a thought in mind that in 20+ years ...