i tried ncbi but i don't the steps how i can start?i found fasta of my gene but how i could deign cDNA with double strand and associated with cDNA primes forw/backw ???
Using protein sequence data, you can predict putative sequences for each aminoacid. If you are able to find mRNA seq data of GOI, you can design as like below;
- fasta seq is in 5-3 direction. end of your seq will be one of your primers.
From beginning of you seq your primer will be like that:
Seq: ATTCCTAG
Pr: CTAGGAT
BUT if you're planning to use it for clonning purpose, add 1-2 restriction sites additionally to end of your primer.