Can anyone tell me why a Taqman PCR amplicon should be restricted to 50-150 bps? What happens if the product is aroudn 200 bps. I am in a tight spot in my sequence and can only do a 200 bp amplification. Any suggestions would be appreciated.
With larger amplicons you are likely to have unconsistent results. This system works on the base of fast amplification and large amplicons get amplified slower (obvious). The IDEAL is an amplicon between 50-150 bp, but you can go as high as 400 bp for Taqman. So, 200 bp is not going to be a huge problem, just make sure you run samples in triplicate (just in case).