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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.

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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)

Cis or trans? That's what matters.
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