Primer Design for nucleotide sequence - (Aug/27/2008 )
Could any one help me in how to perform primer design for a nucleotide sequence???
i attach a PIc for a nucleotide sequence in order to get F and R primers for PCR
You have to know where your primers should bind. If you have some room you can check different primer sequences and use the best one. Use netprimer to check for the quality and Tm:
- You should have several G or C and the 3' end so that Pol doesn't detach. Hence, at the 5' end should be only a few GC's
- Primer should be between 15-21 nucleotides long (depends on the Tm that you need). Tm around 60 should be OK so that the annealing temp in your PCR reaction will be approx. 55