Any suggestions on primers design for 5' RACE PCR?

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Tatiana Kapelinskaya

Dear Britt, you're right, the primer should be antisense (revers complement) regarding the mRNA sequence and GSP1 and GSP2 need to be in the same orientation.They are used to reduce the occurrence of non-specific PCR products. I advise you to read the manual of the company Clontech.

Britt Merlaen

so GSP1 and GSP2 also need to be antisense regarding the mRNA sequence?

Of course yes! Try to paint the letters (ATGC) on both chains and will understand all . Don't forget to mark on the each of chain 5' and 3' ends.

Thanks a lot!