Questions tagged [ligation]

Is the process of covalently link two DNA ends. The process is usually done by the enzyme class of ligases.

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What is the status of the cfDNA ends?

I am working on a protocol to ligate cfDNA, extracted from blood, into some vector. I have looked into the literature but I cannot find any detail about the status of the ends of the cfDNA. Are they ...
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Ligation without purifying insert

I am planning to insert a 45 bp sequence in a vector. After restricting my insert to create compatible sticky ends, I am finding no way to clean it up. Is there any way for cleaning this fragment ...
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Is blunt end repair and A-tailing necessary when adaptors can be custom made?

I'm planning on creating a RADseq library, which naturally involves digesting genomic DNA following P1 adaptor ligation. I've read a few protocols that require any over-hangs resulting from ...
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How are split inteins joined?

Do they form a peptide bond? Or is it just some affinity/van der Waals interaction? Is the mechanism enzyme catalized? Are the residues/moieties at the join site known? Could they be introduced to ...
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3' or 5' DNA capping protein

Does anyone know of a protein/molecule that can be attached to one end of the DNAs temporarily ? eg. i would like to only to allow one strand of DNA be ligated to another DNA molecule to either it's 3'...
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How to ligate DNA to an RNA molecule

Is it possible to ligate a small DNA molecule to mRNA? I was reading a protocol from Current Protocols in Molecular Biology and one of the procedures involve ligating a DNA molecule to mRNA using a ...
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Why are ligases added with the target sequence in sequencing by hybridisation (SBH)?

I understand that labelled n-mer probes in arrays hybridize with complementary sequences in the target DNA, but I don't know what the role of ligase is in the SBH process. Is it involved in the ...
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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 ...