PCR buffer with Tris-HCl and Ammonium sulfate (4:1)-does it exist?
junk at hgmp.mrc.ac.uk
Thu Feb 5 13:21:50 EST 2004
Historians believe that in newspost
<6b414df1.0402050504.3a95d575 at posting.google.com> on Thu, 5 Feb 2004,
Elisabet W <elisabet.witso at chello.no> penned the following literary
>Can you by a PCR buffer which contains Ammonium sulfate together with
>Tris-HCL (pH 8.5), when the consentration is 4:1?
I would not advise using a PCR buffer with more than 20mM AmmSO4 in the
1x mix so that would give a Tris of only 5mM. IMHO, that is
insufficient buffering capacity and I would not run below 10mM Tris.
I have seen and used a variety of buffers from 10mM Tris + 20mM AmmSO4
up to 75mM Tris + 20mM AmmSO4. pH from 8.3 to 9.0
>Or do I have to make these reagents by myself?
Straight forward. Trizma preset from Sigma plus AmmSO4. Autoclave at no
more than a 10x stock with or without Mg. If using high Tris plus
AmmSO4, beware of pptation over time in the buffer. It will re-dissolve
when heated but can be disconcerting when one's previously clear buffer
I love deadlines. I especially like the whooshing noise they make as
they go flying by.
More information about the Methods