Re: 谁有sequence cosmid的经验？# Biology - 生物学
c*t
1 楼
I don't have the protocol w/ me, but I believe that you can look
it up in the journal Biotechniques. It is a quite reliable method
which is basically:
1. get a set of degenerate primers, usually designed specifically
for your organism.
2. design a specific primer which binds to one end of your DNA.
3. do 3 rounds of PCR, 3 minute elongation time, 10 minute extention time.
i) very low anealing temperature (35C?) for 25 cycles. take 1 ul,
do a 1:20 (or 1:100?) dilution and
it up in the journal Biotechniques. It is a quite reliable method
which is basically:
1. get a set of degenerate primers, usually designed specifically
for your organism.
2. design a specific primer which binds to one end of your DNA.
3. do 3 rounds of PCR, 3 minute elongation time, 10 minute extention time.
i) very low anealing temperature (35C?) for 25 cycles. take 1 ul,
do a 1:20 (or 1:100?) dilution and