Silver stain protocol

C. S. Prakash Prakash at ACD.TUSK.EDU
Tue Jan 23 13:50:28 EST 1996


Marc:  Here is the protocol we follow for silver staining DNA in GeneAmp Gel=
s.
Procedure modified from Bassam et al 1991. Fast and sensitive silver
staining of DNA in polyacrylamide gels. Analytical Biochemistry 196:80-83.

*       Gently shake the gel in 7.5% (v/v) glacial acetic acid for 10 min
at room temp.
*       Rinse the gel in deionized water twice for about 2 min each.
*       Incubate the gel in 10% oxidizer solution (Bio-Rad #161-0444) for
5-10 min.
*       Rinse the gel in water three times for about 5 min each. Use fresh
deionized water each time.
*       Immerse the gel in silver staining solution (100 mg silver nitrate
and 150 =B5L formaldehyde in 100 mL water) for 20 min.
*       Pour out the silver stain solution, and wash the gel quickly with
deionized water.
*       Immerse the gel in an ice-cold developer solution (8=B0C) (3 g sodiu=
m
carbonate, 300 =B5L formaldehyde, and 200 =B5g sodium thiosulfate in 100 mL
water) until optimal image intensity is obtained.
*       Stop the developing process by immersing the gel in 7.5% ice-cold
glacial acetic acid.
*       Air dry the gel and back it with a GelBond plastic film (FMC
BioProducts, Rockland, ME).


---------------------------------------------
C. S. Prakash                     Prakash at Acd.Tusk.Edu
Tuskegee University          Phone (334) 727 8023
School of Agriculture         Fax     (334) 727 8552
Tuskegee, AL 36088. USA

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