Discussion of all aspects of biological molecules, biochemical processes and laboratory procedures in the field.
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In the scientific papers, If get stuck with the way they write that is kind of arbitrary for instance if you have an assay Buffer (10X) that is concentrated and if I dilute 3 ml of that by adding 30 ml of water which will bring the buffer concentration to 25 mM Tris-HCl, Ph 8.0, containing 140 mM NaCl, 3 mM KCl, and 1 mM MgCl2 and 1 mg/ml bovine serum albumin?
Can you please explain in details???
Also with the enzymatic reaction I gut stuck with information given for the enzyme for instance I have this assignment: an enzyme has is provided in a concentration of mg/ml, activity mU/ml and specific activity of pmol/min/mg that is the one unit required to give pmol of product per minute in buffer condition containing 40 uM of substrate and 100 uM of co-substrate. Can you please help to understand this? And how this is relate to the design of the conditions for the enzymatic assay.
In the scientific papers or in protocol procedure, sometime do not understand the way they write their experimental data. For instance, a Buffer that consists of a multible reagents and labeled (10X). I was asked to dilute it by taking 3 ml of that 10x buffer and add 30 ml of distelled water and I was told that this will bring the buffer concentration 1X
With final concentrations of 25 mM Tris-HCl, Ph 8.0, containing 150 mM NaCl, 3 mM KCl, and 1 mM MgCl2 and 2 mg/ml bovine serum albumin?
Can you please explain in details??? How the buffer was made in detail initially and then how it was diluted from 10X to 1X.
To make it 1x you need to dilute it 10-times, thus for 3 ml you need to add 27 ml, not 30. Or better fill it up to 30 ml. That's a difference.
The way to prepare such buffer is simply prepare buffer with 10-times higher concentrations, i.e. 250 mM Tris, pH 8.0, 1.5 M NaCl, 30 mM KCl, 10 mM MgCl2 and 20 mg/ml BSA.
Cis or trans? That's what matters.
Thanks for this explanation, but I still need your help! How to prepare multible reagent buffer how do I calculate and prepare the concentrated buffer that is 10-times higher concentrations of 250 mM Tris, pH 8.0, 1.5 M NaCl, 30 mM KCl, 10 mM MgCl2 and 20 mg/ml BSA. Let's say my final volume is 500 ml. It is very confusing for me.
8 posts • Page 1 of 1
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