Mastermixes for capillary PCR

nvahsen at hot.hgmp.mrc.ac.uk nvahsen at hot.hgmp.mrc.ac.uk
Fri Feb 25 13:05:42 EST 2000


Hi Lars,

we always make our own mastermix, most important is to include BSA. We
have tried LifeTec/Gibco native Taq and Roche/BM Taq in our mix: It's
the same performance. Also the same for comparison DNA hyb master or
DNA SYBR green master with our own mix. It's about the same.
You find our mixture published in:

1	N. von Ahsen, E. Schütz, V. W. Armstrong, and M. Oellerich.
Rapid detection of prothrombotic mutations of prothrombin (G20210A),
factor V (G1691A) and methylenetetrahydrofolate reductase (C677T) by
real time fluorescence PCR with the LightCycler. Clin Chem 45
(5):694-696, 1999.
2	N. von Ahsen, M. Oellerich, V. W. Armstrong, and E. Schütz.
Application of a thermodynamic nearest-neighbor model to estimate
nucleic acid stability and optimize probe design: Prediction of
melting points of different mutations of apolipoprotein B 3500 and
factor V Leiden with a hybridization probe genotyping assay on the
LightCycler. Clin Chem 45:2094-2101, 1999.
3	N. von Ahsen, M. Oellerich, and E. Schütz. Using two reporter
dyes without interference in a single tube rapid cycle PCR:
alpha1-antitrypsin genotyping by multiplex real time fluorescence PCR
with the LightCycler. Clin.Chem. 46 (2):156-161, 2000.

There is also info about how to prepare your own mix on the idahotec
homepage (if Roche has not by now forced them to take it down). This
is the CT Wittwer mix and basically also what we are using.

Regards
Nicolas 



On Fri, 25 Feb 2000 06:14:15 -0800, lors
<lorsNOloSPAM at mcafeemail.com.invalid> wrote:

>Hi everyone,
>I'm testing a Roche LightCycler, and I would like to know if
>anyone has a good recepie for Mastermixes, or if anyone happens
>to know what roche puts in their mixes.
>
>:-) Lars
>
>
>* Sent from RemarQ http://www.remarq.com The Internet's Discussion Network *
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