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Dna ligase

Dna ligase

(Science: enzyme molecular biology) enzyme involved in dna replication. The dna ligase of E. Coli seals nicks in one strand of double stranded dna, a reaction required for linking precursor fragments during discontinuous synthesis on the lagging strand. Nicks are breaks in the phosphodiester linkage that leave a free 3_ OH and 5_ phosphate. The ligase from phage t4 has the additional property of joining two dna molecules having completely base paired ends. Dna ligases are crucial in joining dna molecules and preparing radioactive probes (by nick translation) in recombinant DNA Technology. An enzyme that promotes the joining up of adjacent bonds in a polynucleotide Chain, which is used in [[gene.


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MOLECULAR CLONING

... to mention, that synthesized oligos have 5'-OH and 3'-phosphate ends... DNA ligase removes 3'-phosphates and replaces it with -OH. Thus, synthesized oligos without polynucleotide ...

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by kk
Fri Jul 19, 2013 12:50 pm
 
Forum: Molecular Biology
Topic: MOLECULAR CLONING
Replies: 9
Views: 5575

MOLECULAR CLONING

... oligos need to be modified in any way before the ligation reaction. And yes, they need a 5'-phosphate added, by polynucleotide kinase, so the ligase can actually generate the covalent link.

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by kk
Wed Jul 10, 2013 2:58 pm
 
Forum: Molecular Biology
Topic: MOLECULAR CLONING
Replies: 9
Views: 5575

Inserting cDNA into a retroviral vector?..

... of single stranded RNA. Is it possible to insert double stranded cDNA into this vector? If yes, what are the processes that need to happen in order to make this occur? And can I use DNA ligase to seal the fragments of cDNA and retroviral vector?

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by x555
Sun Mar 10, 2013 1:43 pm
 
Forum: Genetics
Topic: Inserting cDNA into a retroviral vector?..
Replies: 3
Views: 2719

Re:

... to do with the splicing nature of the gene.... he suggested that we stop trying to clone the whole gene and look at the splicing nature of ligase 6 before we re-attempt any cloning or over-expression. In order to investigate the splicing nature of the gene, I created primers for each exon ...

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by polkadotshorts
Mon Dec 10, 2012 7:12 pm
 
Forum: Genetics
Topic: How to clone a gene with lots of splicing intermediates?
Replies: 6
Views: 3386

Re:

... intron-free clone is the most important part. Moving on from that, I am trying to write some content about the alternative splicing nature of the ligase 6 gene from a gel image that I have taken. If you have the time, I would really appreaciate it if you could help me out! I am finding it all ...

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by polkadotshorts
Sat Dec 08, 2012 3:52 pm
 
Forum: Genetics
Topic: How to clone a gene with lots of splicing intermediates?
Replies: 6
Views: 3386
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