1.Fix gel in 50% methanol for greater than 1 hour rinse gel in dH2O 3 times 5 minutes each place back in 50% methanol for 1 hour. 2.Stain gel in 100 mls of stain solution for 30 minutes 3.Rinse gel in dH2O 3 times 5 minutes each. 4.Place gel in Developer until bands are almost as dark as desired,sually 20' - 2 hours. 5.Stop developing and fix stain by placing gel in 50% methanol.Bands may continue to darken in methanol overnight,it's best to photograph gel immediately. 6.Photograph on light box.(for type 57 film use settings of 1/125 sec,f=22; for type 55 film use settings of 1/15 sec,f=11) 7.Wrap gel in prewet BioWrap® and allow to dry overnight Stain solution dissolve 0.8 g of AgNO3 in 4 mls dH2O add a mixture of the following.... 21 mls of 0.36% NaOH plus1.4 mls of concentrated NH4OH (a little extra may be needed; add just enough to get silver to dissolve wait for lag!)then bring up to 100 mls with dH2O (by adding 72.8 mls) Developer 500 mls of dH2O 2.5 mls of 1% Citric Acid (monohydrate!) 0.25 ml of 36% formaldehyde |
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