about flanking sequnce
Posted 12 April 2009 - 04:57 PM
Posted 12 April 2009 - 07:13 PM
I am not sure I get yr problem but if you are trying to ask about the number of bases that you need to keep on either side for the enzyme to act, then goto the NEB site and it will tell you. I generally choose the bases on the basis of sequence of the gene, if it is GC rich then A or T or vice versa, just make sure that you don't engineer a stop codon.
Posted 14 April 2009 - 07:40 PM
I think you just ask company to synthesize two DNA oligos for you:
5' TCGAC...36nt...GAT 3'
5' ATC...complemental reverse 36nt...G 3'
let them anneal, you can ligate to where you want it insert.
Posted 14 April 2009 - 08:13 PM