4 replies to this topic

[blondy11]

I'm wondering how to prepare this buffer (TBE 10x)?

I need to know if final volume of this buffers is 1liter, when i dissolve each component and add 1 liter the volume is bigger than 1liter. When I make buffers how much water i should use? Can I correct the volume of buffer with components  to 1liter  or i should add exactly 1 liter to the components. In second case the volume of buffer will be bigger than 1 liter. It is silly question but ?

[casandra]

blondy11, on 06 April 2012 - 10:58 AM, said:

I'm wondering how to prepare this buffer (TBE 10x)?

I need to know if final volume of this buffers is 1liter, when i dissolve each component and add 1 liter the volume is bigger than 1liter. When I make buffers how much water i should use? Can I correct the volume of buffer with components  to 1liter  or i should add exactly 1 liter to the components. In second case the volume of buffer will be bigger than 1 liter. It is silly question but ?

You have to dissolve in less than the final volume, in your case, if you're preparing 10x TBE stock solution in 1 liter, dissolve all the components first in 700 to 800 ml, then complete to a liter.

[blondy11]

So there is no other way to do it  or will be incorrect?

[casandra]

blondy11, on 07 April 2012 - 08:38 AM, said:

So there is no other way to do it  or will be incorrect?

you're preparing a solution here (a certain amount of solute in a certain volume of solvent) so do you think you will have the same concentration if you dissolve 108 g of Tris base in 1 liter and let's say, in 1.2 liters? If you ended with with a lot more final volume than you initially computed for, then just recompute and readjust for a bigger volume.

[Deepak Anand]

There must be a book by name "Molecular Cloning" by Sambrook. There you will get all the molecular biology reagent preparation and standard protocols. Y need to see the Volume 3 Appendices (Page no A1 onwards)
Here I will tell you
5x stock for 1Liter
54gm Tris base
27.5gm boric acid
dissolve them and add
20 ml of 0.5 M EDTA solution
final volume to 1 liter