Does anybody knows how to interpret Non radioactive EMSA

[DNAIA]

Hi all,

I am attaching my EMSA result I know it is not perfect because it needs more optimization, however, I realized that I have very small shift in lane 5. Also,
I realized that when there is a shift, the shifted band becomes more fainter in color compared with the free RNA band, does anybody has explanation for this?


from right to left:

1- free RNA
2-biotinylated rna+ protein 100nm
3-biot. rna + protein 200nm
4-biot. rna+protein 400nm
5-biot. rna+protein 600 nm

[DNAIA]

Originally Posted by DNAIA

Hi all,

I am attaching my EMSA result I know it is not perfect because it needs more optimization, however, I realized that I have very small shift in lane 5. Also,
I realized that when there is a shift, the shifted band becomes more fainter in color compared with the free RNA band, does anybody has explanation for this?


from right to left:

1- free RNA
2-biotinylated rna+ protein 100nm
3-biot. rna + protein 200nm
4-biot. rna+protein 400nm
5-biot. rna+protein 600 nm

folder

[mmorgan]

Which way round is your film?

[blaszcz]

Form its look like RNA-protein complex migrates faster than the free probe. It means that RNA in complex with protein has quiet diferent structure than alone. If the orientation of the gel is correct than my explanation is like above.

Best