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PCR with one primer? is that possible?

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PCR with one primer? is that possible?

Postby kyleenyeoh » Fri Feb 05, 2010 4:38 am

hie everyone, im a student currently taking genetic course, i have to deal with lots of PCR stuffs. Recently, i got completed cDNAs from RACE PCR from my senior, i will like to amplify the full lengh of 3' end of my gene of interested. I wonder if i could use only one gene specific primer(forward primer) for my desire gene without a reverse primer and run the PCR? thanks very much for da response. =)
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Postby JackBean » Fri Feb 05, 2010 7:30 am

There are plenty of PCR applications using only one primer. E.g. your RACE PCR, sequencing or first strand cDNA synthesis
But of course, you will get ony linear increas of amplicone ;)
http://www.biolib.cz/en/main/

Cis or trans? That's what matters.
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Re: PCR with one primer? is that possible?

Postby kyleenyeoh » Fri Feb 05, 2010 2:36 pm

uhhuh... just tat im so confused.. i did the PCR last time, when i ran electrophoresis, i got a band, but da band is just too small in size (i expect it to be much bigger since its 3' full lengh cDNA amplified), i showed it to my supervisor, he said he is not sure what da band is, he told me running PCR with one primer is not working, because every cycle da newly formed DNA will have extra few bases, is it really so? well..because my template is full lengh cDNA, so he adviced me to run PCR again with the oligo dT as my reverse primer.. >.< awwwwwwwww..
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Postby JackBean » Fri Feb 05, 2010 3:26 pm

If that was true, even usual PCR wouldn't work. The point is, that when you use only one primer, every cycle you amplify only that number of new strands, as you had originally in the tube ;)
But if you have the possibility to use both primers, that would be probably better ;)
http://www.biolib.cz/en/main/

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Postby kyleenyeoh » Sun Feb 07, 2010 12:58 am

okies i got it~ ^^ thanks a lot for da help jack =)
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