2 replies to this topic

[science-lover]

Why when we make Forward primer we keep the sequence as it is without doing it in complementary, while we do reverse complement in the reverse primers. any clue

[lab_microbe]

science-lover, on 03 January 2011 - 05:59 PM, said:

Why when we make Forward primer we keep the sequence as it is without doing it in complementary, while we do reverse complement in the reverse primers. any clue

The DNA-polymerase can only sinthesize in  5' to 3' direction. Your fwd primer is in on the beginning  of the + strand in 5' to 3' direction, and your reverse primer is on the beginnig of the -strand in 5' to 3' (or if it easier to visualize for you: on end of the - strand in 3' to 5' direction). So after you amplify your desired  gene with this primers you will get the dsDNA with fwd and reverse primer (http://lslab.lscore....dria/primer.htm).
Also, there are lots of  other useful sites about primer design, just google it. Here are some I came across: PCR_Primer Design, primer_design


Cheers 

Edited by lab_microbe, 04 January 2011 - 04:53 AM.

[science-lover]

Thanks Lab_microbe. this was very helpful.