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Ligase

An enzyme which is capable of binding two smaller components into one single structure.ligase

(Science: enzyme molecular biology) major class of enzymes that catalyse the linking together of two molecules (category 6 in the I.U.B.C.E. Nomenclature) for example dna ligases that link two fragments of dna by forming a phosphodiester bond.


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MOLECULAR CLONING

Not 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 kinase treatment will participate in the ligation reaction with both their ends ...

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by kk
Fri Jul 19, 2013 12:50 pm
 
Forum: Molecular Biology
Topic: MOLECULAR CLONING
Replies: 9
Views: 4108

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: 4108

Inserting cDNA into a retroviral vector?..

... 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: 2149

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: 2507

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: 2507
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