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Electrophoretic Mobility Shift Assay Forum EMSA Forum. Discuss the Electrophoretic Mobility Shift Assay protocols or technique and the assessment of DNA and RNA binding proteins here. Also post questions and troubleshooting about the shift assay.


use RNA on LightShift DNA EMSA Kit

use RNA on LightShift DNA EMSA Kit - Electrophoretic Mobility Shift Assay Forum

use RNA on LightShift DNA EMSA Kit - EMSA Forum. Discuss the Electrophoretic Mobility Shift Assay protocols or technique and the assessment of DNA and RNA binding proteins here. Also post questions and troubleshooting about the shift assay.


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Old 10-28-2010, 04:02 PM
Pipette Filler
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Default use RNA on LightShift DNA EMSA Kit



Hi everybody,

I have in my lab a LightShift EMSA Kit from Pierce designed for DNA binding. I was wondering if I can use RNA instead of DNA with it.

I compared the manual to RNA EMSA one and the only difference seem to be the Binding buffer:

DNA kit: 10X Binding Buffer, 1 ml, 100 mM Tris, 500 mM KCl, 10 mM DTT; pH 7.5

RNA kit: REMSA Binding Buffer (10X), 1mL, 100mM HEPES (7.3), 200mM KCl, 10mM MgCl2, 10mM DTT

Has anybody experience with that?
Do you think I can try to use the DNA buffer, or I should make up the RNA one (I scare RNase doing that)?

Many Thanks

Mingu
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Old 12-09-2010, 10:15 AM
Pipette Filler
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Default Re: use RNA on LightShift DNA EMSA Kit

Well, after trying I can answer myself: in my case both buffers work well. The only thing you don't find in DNA kit are tRNA to use as nonspecific competitor. But you can buy it separately or use Heparin as an alternative.
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