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Plaue assay

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Gateway technology

From what I understand, the gateway technology is a lab method to obtain cloning and expression vectors based on the property of phages to recombine its genetic material with bacterias. However, why do I need to perform the BP reaction, if I want to obtain an expression clone of a certain gene? Why ...

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by miibay
Tue Aug 19, 2014 10:47 pm
 
Forum: Molecular Biology
Topic: Gateway technology
Replies: 1
Views: 214

Re: Gene cloning question

I want to clone a human gene in pET-21a, and transfer to E.coli. I clone the gene in Nde 1 site that it’s sequence is CA/TATG, My question, is The ATG of Nde1 act as a start codon in result I must delete The ATG of my sequence or I can use the ATG of vector in Nde1 site as a start codon? (NOTE: My g...

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by mohajeri
Thu Feb 06, 2014 10:01 pm
 
Forum: Molecular Biology
Topic: Gene cloning question
Replies: 20
Views: 17547

Gene Cloning

Since a PCR product can insert in both direction in the PTZ57 vector and both of clones can grow in a ampicillin LB plates, How can we find the clone that has the correct direction for expression of PCR product?

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by ghaemi100
Fri Jun 21, 2013 9:39 am
 
Forum: Molecular Biology
Topic: Gene Cloning
Replies: 3
Views: 1368

Gene patents may hamper innovations in patient care

“Gene patents are discoveries, not inventions and patenting discoveries may hamper future innovations in developing cost-effective patient care products and services. Scientific and technological innovations in molecular cloning, sequencing, PCR, bioinformatics, biochemical methods etc., have create...

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by TMJohnson
Mon Dec 17, 2012 11:38 am
 
Forum: General Discussion
Topic: Gene patents may hamper innovations in patient care
Replies: 0
Views: 1160

Re:

that would suggest there is some small alternative-intron in your exon, which can be thus split why are you using primers only for one exon at a time? Is your aim to clone all variants or are you fine with only some? Well, we initially had 4 sets of primers. I was going to amplify 4 overlapping reg...

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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: 3239
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