microRNA quantification

3 posts / 0 new
Last post
raihana
raihana's picture
microRNA quantification

 I  have designed RT primers to amplify the mature miR 206  and miR 16 by end point pcr ( from  human colon tissue)  and run on 15 %PAGE followed by silver staining. I am getting multiple bands on the gel in miR 206 lane and not in miR 16 lane.
 Can anybody please tell me what can be the reasons behind this, can these be the non specific products or some isoforms.

Ivan Delgado
Ivan Delgado's picture
There are a number of reasons

There are a number of reasons why this could happen. Assuming that the primers you are using have been validated and optimized using controls, it is possible that your template is degraded or of not good enough quality. One thing you can do is purify a new template and try again. 

If the assay has not been optimized you may want to run an assay that you know works well first and see if the problem is with the primers you are using. 

Good luck.

raihana
raihana's picture
 thanks....hope it works

 thanks....hope it works