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need help in primer design

need help in primer design - PCR - Polymerase Chain Reaction Forum

need help in primer design - PCR - Polymerase Chain Reaction Forum. Discuss and ask questions about PCR troubleshooting, PCR protocols and methods, PCR products, and PCR theory.


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Old 07-27-2011, 08:07 AM
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Default need help in primer design



Hye Everyone!
m working on GABRD gene in humans, accession number (NG_008168.1).... i neeed to amplify all of its exons for analysis.... i've tried alot in primer designing but didnt get just 1 single hit in blast.... i got just 1 hit with in-silico pcr at UCSC but wen i blast those primers they bind on other positions too in human genome.... i m toooo confused, how to design primers which can bind with only my required position.
plz plz plzzzzzzzzzzz help me
thanx
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Old 07-29-2011, 10:51 PM
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Default Re: need help in primer design

If you want to amplify all the exons i suggest than you find the sequence of cDNA, search on NCBI with the name of the gene of interesting, with the sequence of RNA messenger transformed ind DNA dont will have introns in the sequence.
Later you can designing the primers of the start to end of sequence of the gene of interesting
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Old 08-13-2011, 10:34 AM
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Default Re: need help in primer design

Now you can use NEB primer design tool. Here you can choose length and it can automatically cross check non specific amplification other than ur gene of interest

Just type NEB Primer designing tool in google you will get the link

or
you can download
from trial version of NIT vector software and design primer
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