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Primer design


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#1 Debbie Klestadt

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Posted 16 March 2002 - 07:51 AM

Could anyone give me tips on designing my primers for RT-PCR? I will be studying the expression of various cytokines, NOS and membrane receptors from the monocytic cell line THP-1. I intend using Tri Reagent (Sigma) for RNA extraction, then proceed with reverse transcription and finish with with my PCR.

#2 anonymous

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Posted 16 March 2002 - 09:56 PM

For RT-PCR primer design, I usually follow these lines:
1) pcr product site 200-400bp
2) span a intron within amplified region
3) primer length: 18-24mer
4) non complementary between two primers
5) non hair-pin within primeer
6) use some software like MacVector(for Mac), Omega(for PC), and others (there are many)

Hope it helps






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