I have been trying to pick up primers by using CODEHOP. I thought reverse primers could be given raw that I should make reverse-compliment since I could not have the target gene after cloning. Codehop gives me Forward as CCAGAACGACACCGAGgarganttycc -3' and reverse as the compliment (It says) atrctyttrgtGGTCTTGCTGTGGCTCC -5'. Do you know this Reverse primers already complimentary (reverse-compliment) or I need to convert them in both reverse and compliment?
http://blocks.fhcrc.org/codehop.html
Hilal,
if you have the reverse primer in 3'-5' direction (atrctyttrgtGGTCTTGCTGTGGCTCC -5'), you must re-write it in your order in direction 5'-3' like your forward primer.
Hi Alex,
Yes I do it as 5'-3' because it is necessary for ordering. But do you know Should I convert also like 5' GGAGCCACAGCAAGACCac......3' or i need just do it like 5' CCTCGGTGTCGTTCTGGtgrttytcrta 3' ?
atrctyttrgtGGTCTTGCTGTGGCTCC -5' is on reverse strand. It is already complementary sequence to plus strand of DNA.
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