We don't have a Stratalinker or any UV crosslinkers in our lab or our department. Is is absolutely necessary to crosslink RNA while doing a Northern blot? I am planning to do a nonisotopic northern blot. Does this lack of crosslinking affect my results in any way? Also, can I use any of the following as a makeshift UV incubator:
- handheld UV lamp (UVGL-25 from UVP),
- UVP imager with a UV light box (to take agarose gel images DNA/RNA),
- UV tubelight in our laminar airflow hood (to decontaminate the area)
Alejandro Martin
I have not done Northern blots in a long time, but I do remember that Hybond-N+ (from Amersham, now GE) could cross-link RNA simply by putting the filter, RNA side up, for a short time on a pad moistened with low concentration NaOH (I think it was 10 to 15 mM but take it with a grain of salt). Perhaps there is some alternative protocol as well in the booklet for your membrane?
2 votes
1 thanks
- Badges
- Science method
More Shruthi Gobbooru's questions See All
Similar questions and discussions