Re: [DIYbio] visualizing small RNAs

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.

On Feb 10, 2017 3:23 AM, "Mega [Andreas Stuermer]" <masters...@gmail.com> wrote:

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 ;)

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