Re: Did I get correct cDNA clone?# Biology - 生物学
b*r
1 楼
how
1.extract the total mRNA
2.use restriction enzyme to cut out the desired gene
3.design a primer and do a PCR
4.Sequence the DNA and compare with those published sequences to see whether
it encodes the kind of enzyme you want.
5.Southern Blot to see whether pure and whether it is there.
6.Express the cDNA in Ecoli and check protein activity.
7.translate the sequenced cDNA to see whether you got the correct protein.
8.Sequence the expressed protein and back translate part of the AA's that has