How to Prepare Digestion Buffer - (Feb/07/2012 )
Hello guys.. I need your help.. I'm going to prepare digestion buffer before DNA extraction. I'm working with cancer cell line.
This is the list of reagents:
100 mM NaCI
10 mM Tris.CI, pH 8.0
25 mM EDTA, pH 8.0
0.5 % SDS
0.1 mg/ml proteinase
* the final volume should be 1 ml.. can anyone show me the correct steps to prepare each of the materials?
i really appreciate your kindness... Thank you very much.
I would prepare a stock solution of your buffer and reagents. Make solutions of 1 M Tris-HCl pH 8.0, and 0.5 M EDTA pH8, autoclave them; 20 % SDS. Then you can prepare your basic buffer, e.g. 100 ml: 0.58 g NaCl, 1 ml Tris-solution, 5 ml EDTA solution, 2.5 ml SDS, fill up with water.
When preparing the digestion add fresh proteinase to the desired volume.