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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8
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2answers
252 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 ...
0
votes
1answer
22 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 ...
3
votes
0answers
60 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
votes
0answers
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: ...
2
votes
0answers
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 ...
2
votes
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 ...
2
votes
0answers
307 views

Can Pfx polymerase add only one 3' A overhang?

I am trying to clone a PCR product that was amplified using Pfx polymerase into pGemT vector. I had to A-tail the PCR product using Taq polymerase since Pfx only generates blunt end products. My ...
1
vote
0answers
26 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 ...
1
vote
0answers
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 ...
1
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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 ...
1
vote
0answers
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 ...
1
vote
0answers
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. ...
1
vote
0answers
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 ...
0
votes
0answers
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 ...
0
votes
0answers
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 ...
0
votes
0answers
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 ? ...
0
votes
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?