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Ligate

Ligate

to tie. As, for example, the surgeon ligated the artery.


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Producing unknown protein

... to the expression system (you have a choice of bacterial or cell free expression systems) 3. Synthesize sequence (IDT can do it for you). 4. Ligate into vector (vector depends on the expression system)/clone vector. 3. Transform and/or express ...

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by Cat
Mon May 20, 2013 1:56 pm
 
Forum: Microbiology
Topic: Producing unknown protein
Replies: 2
Views: 2004

MOLECULAR CLONING

Hello, I have a related question. I try to ligate annealed oligos into double digested (different enzymes) plasmid. The oligos I annealed were not modified with phosphates. Do I need to phosphorylate or dephosphorylate any of the two? As far as ...

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by kk
Tue Feb 12, 2013 2:48 pm
 
Forum: Molecular Biology
Topic: MOLECULAR CLONING
Replies: 9
Views: 5208

Re:

... to amplify 4 overlapping regions of the gene, run them on a gel, cut out the bands of expected size, purify them, and eventually sequence and ligate them together to generate the full length gene which I would then clone and amplify. However, there were troubles with the isolation of the bands. ...

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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: 3192

Cloning/Transformation question

also sometimes when i ligate short oligos, i use different inster:vector ratios into different plates just to see the pattern.. hope it helps too!

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by bravebeaker
Sat Dec 01, 2012 7:00 am
 
Forum: Molecular Biology
Topic: Cloning/Transformation question
Replies: 4
Views: 2260

Cloning/Transformation question

... into some chemically competent E. coli cells. I was wondering what I should run as a negative control? I know if the cloning works the plasmid ligates and an antibiotic resistant gene on the plasmid starts to be expressed. If E. coli cells take up the plasmid they will obviously grow on the ...

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by coppcw08
Fri Nov 30, 2012 12:24 am
 
Forum: Molecular Biology
Topic: Cloning/Transformation question
Replies: 4
Views: 2260
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