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mature form of a gene via PCR

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mature form of a gene via PCR

Postby biology_06er » Mon Aug 15, 2011 11:35 am

Hey guys,

So I have recently PCRed my complete gene of interest/carried out blue white selection and the next step (well not next step but something I will have to do very soon) is to PCR the mature form of the gene..so what is the general process here..from genomic DNA, I used primers etc and got a lovely band when I ran my gel but if I only want to clone the mature from what will I have to do...(I assume this means I have to have the signal peptide removed)...will I need diff primers or something..hope I make sense.

cheers
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Postby uday21 » Wed Aug 17, 2011 11:54 am

I think to get mature form of Gene of your interested. you have to use cDNA of your gene not the genomic DNA to do the pcr.
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Postby JackBean » Wed Nov 02, 2011 5:50 pm

that depends, what exactly you mean by mature gene. Whether after splicing (then use cDNA) or mature protein without signal peptide (use cDNA with primers after the signal peptide)
http://www.biolib.cz/en/main/

Cis or trans? That's what matters.
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