{Q} Effective Inverse PCR?

Roland Saldanha rsaldanh at mail.utexas.edu
Wed Aug 2 20:39:19 EST 2000


In article <020820001614466447%dmicklem at cmgm.nospam.invalid>, David
Micklem <dmicklem at cmgm.nospam.invalid> wrote:

> In article <8m8usc$hs2$1 at nnrp1.deja.com>, Andreas Firzinger
> <a_firzinger at my-deja.com> wrote:


>  and 4units
> >T4 ligase/µl, this means 300 units T4 ligase in one reaction!!



> The Drosophila genome project protocol at
> 
> <http://www.fruitfly.org/about/methods/inverse.pcr.html>
> 
> only uses 2 weiss units/reaction.

> David
> 


I am unfamiliar with the Biotechniques paper but they may be using
Cohesive End Ligation Units (used by several manufacturers like NEB and
defined as giving 50% ligation of HindIII digest of lambda in 30 min at 16
C). One Weiss Unit = 67 Cohesive End Ligation units according to the NEB
catalog.  Hence you are using about 137 cohesive end units. 


Roland






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