Hey Mega,
I work with small RNAs as a major part of my dayjob, and I can tell you from first-hand experience that agarose will not be able to provide the kind of resolution you're looking for. You'll need to be using a high-percentage denaturing PAGE gel - preferable 15-20% TBE-Urea.
You can minimize your exposure either by using pre-cast gels - if you have access to a compatible rig (Bio-Rad, Invitrogen, possibly others - look around on eBay for a used gel rig) - or pre-mixed reagents for casting your own. The second option obviously requires a lot more gear, and casting PAGE gels is quite a bit more finicky than pouring agarose gels. Either way, keep in mind you'll need a denaturing loading buffer, a way to stain your RNA (e.g. SYBR Green II), and a way to safely dispose of everything afterwards.
I can dig up some manuals / protocols at work tomorrow if you need more info.
Hi, I was wondering if anyone has worked with small RNAs (20-50 bp) in a DIY setting. I heard you can't visualize them on an Agarose gel well. No chance you can distunguish 22 bp and 20 bp?
Ideally some method without toxic ingredients ;)
--
-- You received this message because you are subscribed to the Google Groups DIYbio group. To post to this group, send email to diybio@googlegroups.com. To unsubscribe from this group, send email to diybio+unsubscribe@googlegroups.com . For more options, visit this group at https://groups.google.com/d/forum/diybio?hl=en
Learn more at www.diybio.org
---
You received this message because you are subscribed to the Google Groups "DIYbio" group.
To unsubscribe from this group and stop receiving emails from it, send an email to diybio+unsubscribe@googlegroups.com .
To post to this group, send email to diybio@googlegroups.com.
Visit this group at https://groups.google.com/group/diybio .
To view this discussion on the web visit https://groups.google.com/d/msgid/diybio/3290317f-60f1- .4b85-b6e2-05317c9216cb% 40googlegroups.com
For more options, visit https://groups.google.com/d/optout .
-- You received this message because you are subscribed to the Google Groups DIYbio group. To post to this group, send email to diybio@googlegroups.com. To unsubscribe from this group, send email to diybio+unsubscribe@googlegroups.com. For more options, visit this group at https://groups.google.com/d/forum/diybio?hl=en
Learn more at www.diybio.org
---
You received this message because you are subscribed to the Google Groups "DIYbio" group.
To unsubscribe from this group and stop receiving emails from it, send an email to diybio+unsubscribe@googlegroups.com.
To post to this group, send email to diybio@googlegroups.com.
Visit this group at https://groups.google.com/group/diybio.
To view this discussion on the web visit https://groups.google.com/d/msgid/diybio/CALZhpzJfeOuSt_ZUddb26f-GR-i9FXxxP5ecLfJgK%2BbK3M9_Bg%40mail.gmail.com.
For more options, visit https://groups.google.com/d/optout.
0 comments:
Post a Comment