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Ligases

Ligases

(Science: enzyme) a class of enzymes that catalyze the formation of a bond between two substrate molecules, coupled with the hydrolysis of a pyrophosphate bond in atp or a similar energy donor.

registry number: EC 6.


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ligation problem although try everything,suggestions?

... in primers, it could lead to problems with inefficient digestion as well. These are just two but possibilities are many. Since you tried several ligases, problem might be not with them but at some prior step.

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by dustman
Thu May 24, 2012 10:42 am
 
Forum: Molecular Biology
Topic: ligation problem although try everything,suggestions?
Replies: 7
Views: 4091

Re: ligation problem although try everything,suggestions?

I haven't done any lab work for many years, but in the old days, the buffers that came with ligases would often form a precipitate and needed to be vigorously vortexed before using. Have a look at your buffers to see if there's any precipitates. If so, try vortexing the ...

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by wbla3335
Wed May 16, 2012 5:14 pm
 
Forum: Molecular Biology
Topic: ligation problem although try everything,suggestions?
Replies: 7
Views: 4091

Genetic Eng: How do you ID the gene from the protein

... I've googled this up and down the yin-yang and looked in various text books (e.g. Lodish et al.). Loads of info about restriction enzymes and ligases and vectors, which I know about. They all jump over the interesting bit at the beginning and simply say, isolate the gene, next... What comes ...

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by nimrod1212
Sun Jun 05, 2011 6:15 am
 
Forum: Molecular Biology
Topic: Genetic Eng: How do you ID the gene from the protein
Replies: 3
Views: 1682

Proteins in Mitosos and the Cell Cycle :)

APC/C and SCF are the ubiquitin-protein ligases for the cell cycle. They themselves are not phosphorylated though. Sic1 is attached to S-phase cyclin/CDK, and when it becomes phosphorylated by the next phase cyclin/CDK, then it is marked for ...

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by kolean
Mon Nov 30, 2009 3:10 pm
 
Forum: Cell Biology
Topic: Proteins in Mitosos and the Cell Cycle :)
Replies: 4
Views: 3122

DNA ligases

After you "cut" DNA fragments using restriction enzymes, and then are "pasting" them together using DNA ligase... do you need to select a specific DNA ligase that matches with the specific restriction enzyme that was used?

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by SysBio
Fri Mar 27, 2009 3:40 am
 
Forum: Molecular Biology
Topic: DNA ligases
Replies: 1
Views: 1849
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