Hi, I am experiencing signal on the well position. Notes 12...

SC
Sung Ki Cho
Aug 27, 2014
protocol Protocol: Analysis of RNA-protein Interactions Using Electrophoretic Mobility Shift Assay (Gel Shift Assay)
Hi, I am experiencing signal on the well position. Notes 12 recommends to use BSA to minimize it, but how about the concentration of BSA to improve the issue? Thank you in advance.

SK
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saiprasad Goud palusa Author Answered Aug 28, 2014

Department of Biology, Colorado State University, USA

Hi,
we normally use 100ug/ml final concentration of BSA for this, but based on your conditions you have to optimize this.
And also your protein should be purified as much as possible
Not exceed more than 15ul of loading volume, if you increase the loading volume, you are increasing the salt concentration and this will effect on PH and finally it will create problem on running condition. If possible add 1 ul tRNA in addition Heparin sulfate.
good luck
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