# Problem in making SDS-page Sample buffer - (Sep/03/2006 )

If i want to make a stock of sample buffer [SDS-page]

Composition:
1. 6% SDS
2. 26.4% glycerol
3. 6% beta-mercaptoethanol
4. 0.19M Tris [pH 6.8]

If i want to make a 10ml sample buffer, then i should
1. USe how many grams of SDS?
2. HOw much (ml) of glycerol?
3. How much (ml) of beta-mercaptoethanol
4. how much (ml) of 0.5M Tris pH 6.8 ?

THanks

-peggybee1-

This recipe tells you how many grams or mls you should use for 100ml of solution, therefore you should use the tenth part of what you have in the recipie. Then you should calculate the dilution factors for beta-mercapto and Tris

-dnafactory-

QUOTE (dnafactory @ Sep 3 2006, 04:10 PM)
This recipe tells you how many grams or mls you should use for 100ml of solution, therefore you should use the tenth part of what you have in the recipie. Then you should calculate the dilution factors for beta-mercapto and Tris

In instance, "6% SDS" means that you need 6g of SDS for a volume 100ml => 600mg of SDS for 10ml.

-behappy736-

QUOTE (peggybee1 @ Sep 3 2006, 06:37 AM)
If i want to make a stock of sample buffer [SDS-page]

Composition:
1. 6% SDS
2. 26.4% glycerol
3. 6% beta-mercaptoethanol
4. 0.19M Tris [pH 6.8]

If i want to make a 10ml sample buffer, then i should
1. USe how many grams of SDS?
2. HOw much (ml) of glycerol?
3. How much (ml) of beta-mercaptoethanol
4. how much (ml) of 0.5M Tris pH 6.8 ?

THanks

6g sds/100ml
1M tris =121.1/1000(Mol.Wt / 1000)
and further u calculate for 0.19M

-fozia-

Hi,

I dont want to b rude, but it would b better for u to calculate it yourself (sort of practice) and post the answers u calculated so that people could verify it. This will help u in future calculations.

others have replied to ur questions.

good luck

-scolix-

it looks as a protocol for double-or 2.5-fold-concentrated; think of a 4- or 5-fold concentrated sample buffer f.i. if using liquid samples as lysats

-The Bearer-

Hmmmm... you know what the original question resembles ... a question one might find on an undergrad biochem lab quiz.

As was pointed out, it would be more beneficial to you to try the answer yourself and post the answers here.

That way you'll get the benefit of understanding how to work through the problem. It really isn't going to help you at all if someone just posts the answer for you.

What would you do the next time you need to change the volume or concentration of one of the components?

-fossiljim-

really there are some people like me ............face e terrible situation for a poor calculating knowledge...........i really feel sympathy for the people like this b'cause i know ...........i understand.........so we should try first...........now i feel may i have some improvent....i can calculate........but truly speaking sometimes some confusion arises.........
best of luck

-T. reesei-