Silver staining - PCR SSCP

ulrike.mau at uni-tuebingen.de ulrike.mau at uni-tuebingen.de
Sun Jan 26 10:06:50 EST 1997


On Thu, 23 Jan 1997 18:14:36 GMT +100,
Schenker at MPIMG-Berlin-Dahlem.MPG.DE (Martin Schenker) wrote:
>1 .FIX:  			10% acetic acid; 20 min
>2. RINSE: 		H2O; 2 min 3 times
>3. IMPREGNATE: 		AgNO3 (1g/l), 1,5ml 37% HCOH/liter, 30 min
>4. RINSE: 		H2O 20s (optional)
>5 .DEVELOP:		Na2CO3 (30g/l), 1,5 ml 37% HCOH/liter, Na2S2O3*5H2O (2mg/l);
>			 2-5 min
>6. STOP:		10% acetic acid: 5 min
>It is fast (ca. 60 min.) reliable and suitable for double and single strand DNA
>Greetings, Martin

Hi Paulo and Martin!
It is not necessary to add HCOH in the impregnation step. It works
fine without it and it saves you money and toxic agents.
You can also reduce the time in the impregnation step from 30 minutes
to 20 minutes (even with 1mm gels).
Another way to save time is to fixate the gel 10 min in 1% HNO3.
If you do not have the time to stain your gels the same day, you can
put them into 10% Ethanol overnight.
We prepare about 500 ml of the AgNO3 solution and use it about 4-5
times. We do this for all solutions except the developing solution.
For the latter we prepare a 3% Na2CO3 solution (about 2 liters). For
using we take about 300 ml and add 50 µl of HCOH per 100 ml. We do not
use Na2SO3. And it works perfectly.
Good luck !
Greetings
Ulrike Mau
(ulrike.mau at uni-tuebingen.de)



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