pcr with NH4+
drm21 at mole.bio.cam.ac.uk
Thu Jun 26 11:48:19 EST 1997
In article <moellere.15.00A2F12D at pz350.ipsp.uni-hohenheim.de>,
moellere at pz350.ipsp.uni-hohenheim.de (EVELYN MOELLER) wrote:
>I would like to know if anyone on the net has experience with including NH4
>ions (additional to MgCl2) into the PCR buffer and knows which concentrations
>I used the Quiagen PCR buffer which contains NH4 ions but the
>company is not willing to inform customers about the concentration used in
>their buffer. I will try different concentrations now, but may be that anyone
>already has done it and can indicate me a useful concentration/range.
>This buffer improves some of my crucial PCRs. But I want to make my own
>buffer instead of buying the expensive Quiagen kit (including the polymerase
>and the Q-solution which I do not need) all the time..... and I think a
>scientist should know what he/she is doing instead of using "undeclared"
>products and supporting this fatalkit philosophy....
>Evely Moeller, University Hohenheim, Inst. Plant Breeding, Germany
>moellere at uni-hohenheim.de
AFAIK, NH4 buffers have been in widespread use for quite a while.
I have recipes for 10x buffers of: 160mM (NH4)2SO4, 670mM Tris pH8.8 at 25,
0.1% Tween20, +MgCl2 to taste.
KlenTaq (Barnes et al, for use in long PCR) came with buffer PC2, which at
500mM Tris pH9.1, 160mM (NH4)2SO4, 25mM MgCl2 and 1.5mg/ml BSA.
This buffer has worked really really well for me for both regular and
If you have a look at recent posts (eg with www.dejanews.com), you may find
mention of betaine as a new wonder-additive, and the references I think.
Does anyone know if this is whats in Qiagen's new buffer.....
D.R.Micklem, Time flies like an arrow...
Wellcome/CRC Institute, Fruit flies like a banana.
Cambridge CB2 1QR, UK Email:drm21 at mole.bio.cam.ac.uk
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