2 replies to this topic

[gayle]

Hi everyone

Could anyone please tell me as to which method,  RT-PCR or Northern Blot analysis is better for detecting alternatively spliced RNA.  Also which kit from which company is preferrable for carrying out northern blot analysis.


Thanks
Gayle

[mikew]

I guess everyone has different experiences with this, but here's mine.
If you have a great probe, Northern blotting is the way to go to examine
alternate splice variants. You will see most (if not all) of the messages that way.
I have tried several Northern protocols, including the "classical" method using Dernharts (sp?), Rapid hybe from Amersham and Zeta probe from Biorad. By far, I got the best results with Zeta probe. Almost no background, easy to master but requires long expposure time. Rapid Hyb is faster, more sensitive, but often gives higher background (in my hands).

[gayle]

mikew, on Dec 23 2004, 11:20 AM, said:

I guess everyone has different experiences with this, but here's mine.
If you have a great probe, Northern blotting is the way to go to examine
alternate splice variants. You will see most (if not all) of the messages that way.
I have tried several Northern protocols, including the "classical" method using Dernharts (sp?), Rapid hybe from Amersham and Zeta probe from Biorad. By far, I got the best results with Zeta probe. Almost no background, easy to master but requires long expposure time. Rapid Hyb is faster, more sensitive, but often gives higher background (in my hands).

Thank you so much for your advice.
how about the northern blot kit from ambion- northernmax kit.