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PCR using CDNA as template - (Aug/29/2011 )

Hello,
I have been trying to amplify a 3'UTR region of an RNA virus. SO i initially made the cDNA and now doing end-point PCR to amplify the UTR region.My primers are to the ends of the UTR region along wiht the restriction sites attached to the prrimers.Total product size 616bp.
I have been getting a clear band in the b-actin control run in the same run, but not the band of my interest. but there are loads of primer dimers at the end!!any suggestions???is it just the annealing temperature?

-Su.J-

What annealing temperature are you using? Have you analysed your primer sequences for dimers and/or hairpin structures? and what is your melting temp of your primers? If not, a great program is oligo analyzer. just punch in your primer sequence and it gives you all the information you need

-SabbathWarPig-

a

-SabbathWarPig-