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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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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: 1006

Questions on the Sodium Potassium Pump and Enzymes

... the enzyme and the substrate working at an active site? For example DNA Ligase forms phosphodiester bonds between Okazaki fragments... What about when one job involves ...

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by JoWillyTso
Mon Nov 05, 2012 5:50 pm
 
Forum: Cell Biology
Topic: Questions on the Sodium Potassium Pump and Enzymes
Replies: 3
Views: 1545

Re: ligation problem although try everything,suggestions?

I've bought the ligase 3 days ago. I was using NEB T4 DNA ligase.Now I'm using Quick ligase. I read at somewhere I can't remember now, it says do not spin ligase. ...

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by genetherapy
Wed May 23, 2012 2:49 pm
 
Forum: Molecular Biology
Topic: ligation problem although try everything,suggestions?
Replies: 7
Views: 2023

ligation problem although try everything,suggestions?

... same enzymes and make sticky(cohesive) end ligation. I used Quick ligase at room temperature(25 degrees) tried 5 minutes as the protocol said (NEB)alternatively try 25 minutes and also tried T4 DNA ligase at room temperature (NEB). I make 1:1, 1:3, 1:5 and 1:10 ligations. ...

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by genetherapy
Wed May 16, 2012 3:31 pm
 
Forum: Molecular Biology
Topic: ligation problem although try everything,suggestions?
Replies: 7
Views: 2023

Inactivate an enzyme

... don't need but must not dephosphorylate your insert, because then the DNA ligase won't be able to ligate the fragments together. It needs the phosphate on the DNA strand.

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by JackBean
Wed Apr 11, 2012 1:14 pm
 
Forum: Molecular Biology
Topic: Inactivate an enzyme
Replies: 7
Views: 1246
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