Concentrating Conditioned Media for use on a non-denaturing agarose gel - (Jan/06/2009 )
Hi all,
I need to concentrate conditioned media to look at RNAs that are secreted. I intend to run the media on a denaturing gel. I'm not sure how to do this without losing small RNAs. Any ideas?
Thank you!
-ciliegia938-
QUOTE (ciliegia938 @ Jan 6 2009, 05:34 PM)
Hi all,
I need to concentrate conditioned media to look at RNAs that are secreted. I intend to run the media on a denaturing gel. I'm not sure how to do this without losing small RNAs. Any ideas?
Thank you!
I need to concentrate conditioned media to look at RNAs that are secreted. I intend to run the media on a denaturing gel. I'm not sure how to do this without losing small RNAs. Any ideas?
Thank you!
Generally you concentrate conditioned media by using Centricon spin columns. But that has cut-offs for proteins and I am not sure how you can judge the size of small RNAs.
The possible solution to me is to do RNA Prep from the conditioned media. Trizol or RNAeasy, although I can see troubles if volume of media is quite huge. May be you can start with low volume RNA prep?
-cellcounter-
QUOTE (cellcounter @ Jan 7 2009, 08:17 AM)
QUOTE (ciliegia938 @ Jan 6 2009, 05:34 PM)
Hi all,
I need to concentrate conditioned media to look at RNAs that are secreted. I intend to run the media on a denaturing gel. I'm not sure how to do this without losing small RNAs. Any ideas?
Thank you!
I need to concentrate conditioned media to look at RNAs that are secreted. I intend to run the media on a denaturing gel. I'm not sure how to do this without losing small RNAs. Any ideas?
Thank you!
Generally you concentrate conditioned media by using Centricon spin columns. But that has cut-offs for proteins and I am not sure how you can judge the size of small RNAs.
The possible solution to me is to do RNA Prep from the conditioned media. Trizol or RNAeasy, although I can see troubles if volume of media is quite huge. May be you can start with low volume RNA prep?
Thanks I am going to try that and see if I can come up with anything else!
-ciliegia938-