miRNA library prep

Vern · 07-21-2011, 08:57 AM

I am having trouble size selecting on the TBE gels for the sm RNA preps? Any advice on what dyes and stains work best?

advanT · 07-25-2011, 03:10 PM

We use SYBR for staining miRNA libraries post PCR. Our buffers gel loading buffers come from a kit. Resolution and separation of miRNAs products works well with SYBR.

shobbir · 03-21-2012, 10:57 AM

what percentage gel works best? did you use a TBE-Urea gel?

NextGenSeq · 03-23-2012, 01:45 PM

8% no urea

shobbir · 03-23-2012, 02:00 PM

i was under the impression it is standard to use denaturing conditions when running RNA?

cascoamarillo · 03-26-2012, 10:33 AM

Hi
Not sure what are you asking for...isolating small RNAs or libraries made from miRNAs?
At least, I did it (small RNA isolation) with 15% denaturing (UreaGel from National Diagnostics) PAGE.
The library purification (eliminate any kind of primer dimer, adapters, etc) I used 10% non-denatuting PAGE.

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07-21-2011, 08:57 AM	#1
Vern Junior Member Location: Virginia Join Date: Jul 2011 Posts: 1	miRNA library prep I am having trouble size selecting on the TBE gels for the sm RNA preps? Any advice on what dyes and stains work best?

07-25-2011, 03:10 PM	#2
advanT Member Location: South Africa Join Date: Oct 2009 Posts: 22	staining TBE gel We use SYBR for staining miRNA libraries post PCR. Our buffers gel loading buffers come from a kit. Resolution and separation of miRNAs products works well with SYBR.

03-21-2012, 10:57 AM	#3
shobbir Junior Member Location: cambridge Join Date: Mar 2012 Posts: 7	what percentage gel works best? did you use a TBE-Urea gel?

03-23-2012, 01:45 PM	#4
NextGenSeq Senior Member Location: USA Join Date: Apr 2009 Posts: 482	8% no urea

03-23-2012, 02:00 PM	#5
shobbir Junior Member Location: cambridge Join Date: Mar 2012 Posts: 7	i was under the impression it is standard to use denaturing conditions when running RNA?

03-26-2012, 10:33 AM	#6
cascoamarillo Senior Member Location: MA Join Date: Oct 2010 Posts: 160	Hi Not sure what are you asking for...isolating small RNAs or libraries made from miRNAs? At least, I did it (small RNA isolation) with 15% denaturing (UreaGel from National Diagnostics) PAGE. The library purification (eliminate any kind of primer dimer, adapters, etc) I used 10% non-denatuting PAGE.