4 replies to this topic

[blasko]

Do you have a second strand cDNA protocol?
Please, share with me.
(There are a loads of first strand kit at manufacturers, but I never find second strand.)


Thanks.

[aj_xy999]

try: http://www.stratagen...ct.aspx?pid=609

[oddie]

I guess no protocol is needed as you obtain first cDNA strand, you can use that first cDNA as template in PCR reaction. Do it as a normal PCR. It works.

[blasko]

I need second strand DNA, because after sythesis I would like to digest by RE.

Thanks stratagene web site.

[cheerioet]

oddie, on Aug 23 2004, 02:10 PM, said:

I guess no protocol is needed as you obtain first cDNA strand, you can use that first cDNA as template in PCR reaction. Do it as a normal PCR. It works.

I am having the same question . I would like to know, if do it like the normal PCR, what type of primer shall I use? I need to have the full length of the cDNA (if possible) to be amplified.

Kits seems too expensive and I am doing for trial. May I know if anyone out there been doing this, and can send me some suggestion and advice?

Thanks in advance.