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MiniProteanThreeRecipe
MiniProteanThreeRecipe

Recipes for the Bio-Rad Mini Protean III SDS-PAGE system

See also the long MiniProteanThreeInstructions without recipes.

The percentage of the resolving gel determines the molecular weights of proteins which can be resolved. 12% is a good generic gel to use and will resolve proteins from 15 kDa to 200 kDa with emphasis on resolving proteins from 15 kDa to 100 kDa. Lower percentage resolving gels will do a better job of resolving larger proteins, but will allow smaller molecular weight (MW) species to migrate off the gel. Higher percentage gels will similarly give extra resolution to lower MW species at a sacrifice of higher MW resolution.

resolving gel
6%
8%
10%
12%
40% acrylamide (29:1 crosslink)
600 μl
800 μl
1.0 ml
1.2 ml
4x resolving buffer pH 8.8
1 ml
1 ml
1 ml
1 ml
H2O
2.4 ml
2.2 ml
2.0 ml
1.8 ml
10% APS
20 μl
20 μl
20 μl
20 μl
TEMED
2 μl
2 μl
2 μl
2 μl
Total volume
4.0 ml
4.0 ml
4.0 ml
4.0 ml

stacking gel
one gel
two gels
40% acrylamide (29:1 crosslink)
117 μl
234 μl
4x stacking buffer pH 6.8
300 μl
600 μl
H2O
783 μl
1566 μl
10% APS
12 μl
24 μl
TEMED
2 μl
4 μl
Total volume
1200 μl
2400 μl

The Mini Protean III needs about 350 ml of running buffer:

Chris, I believe there is aclautly a small amount of weakness in the toxicological defense per Paracelsus. In his time he was speaking of the effects of poisoning as carcinogenic effects were pretty much unknown as a class.Today scientists are divided between the threshold and non-threshold theory camps for carcinogenesis, with the non-thresholders claiming that even a single exposure to the smallest possible immeasurable unit of a substance or condition can cause cancer.There are two problems with this however:1) Many scientists disagree with the theory, claiming that something akin to chaos theory or prinicples of uncertainty enter the equation significantly when the exposures become too small. These scientists claim that a butterfly flapping its wings in Kansas can NOT ever "cause" a typhoon in Vietnam.2) Even if we accept the no-threshold theory the Antismokers' characterization of secondary smoke is misleading. To use the word toxic or dangerous to describe the level of exposure to secondary smoke that would normally be encountered today in any decently ventilated establishment would be like using those words to describe reaching out the door to grab the morning paper without UV safety gloves on or dining in a restaurant where someone was drinking a beverage containing the highly volatile Class A Human Carcinogen ethyl alcohol.Use of "no safe level" to describe those exposures would be highly misleading and destructive.... just as its use to describe secondhand smoke is.Michael J. McFadden??
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