{Q} Effective Inverse PCR?

Andreas Firzinger a_firzinger at my-deja.com
Wed Aug 2 05:58:21 EST 2000


In article <3ath5.860$OQ3.18709 at news2.bora.net>,
  "Hyun Mi Back" <hmback at chollian.net> wrote:
> I try to clone genes using Inverse PCR.  Several times I failed.
Anybody
> could help me with successful methods or strategies?
> Theory is simple, but it does not work yet.
>
> Han
> hbkim at office.hoseo.ac.kr
>
>

I just found two papers in recent BioTechniques (Vol.28, No.5 (2000)).
Never did inverse PCR before, but I'd have to during next three
month. However, what really surprised me was the HUGE amount of
T4 ligase the authors used: 75µl volume with low DNA-concentration
to achieve a high rate of self-ligation (2.5ng/µl) and 4units
T4 ligase/µl, this means 300 units T4 ligase in one reaction!!

Ciao,

Andreas


Sent via Deja.com http://www.deja.com/
Before you buy.






More information about the Methods mailing list