Magnetic Bead-Based RNA Extraction

Use this category for the exchange of ideas, methodologies and references regarding the isolation, manipulation and analysis of RNA. (Extraction protocols, Northern Blot analysis, RNase Protection, Differential Display, In Vitro Transcription, etc.)

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Magnetic Bead-Based RNA Extraction

Postby OneHandedPipetter » Nov 30 2016 11:23 pm

Hi all,

I am working on a Microfluidic Magnetic bead based Assay with an extraction method for RNA. The beads used are coated with a silica coating, and will be extracted using an elution buffer. MY background in my molecular biology only extends to orgo chem separation techniques such as Trizole digest with chloroform or phenol extraction, and precipitation with 100% IPA. I am curious to know if anyone has had any experience with magnetic bead based RNA extraction. I am having trouble determining the appropriate elution buffer pH, does anyone know an optimal pH for silica bead elution. I would think it is similar to a column elution pH. Thank you!

Any feedback I will appreciate!

-C
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Re: Magnetic Bead-Based RNA Extraction

Postby DrShabba » Jul 19 2017 9:10 am

We use a generic elution buffer for bead based methods, but it's basically pH7 TE buffer (pH 7 is best for storage) to my knowledge.

Some people use water for RNA :roll:

With bead based methods, you usually co-precipitate DNA and RNA unlike with traditional methods (or some spin column methods).

Work flow will be something like:
1. Lysis with chaotropic salts (gaunidine +/-SDS) and Proteinase K digest.
(DNASE/RNASE digest can be performed here)
2.Binding to beads, usually with Isopropyl Alcohol binding buffer.
3. 1-2 wash steps, EtOH based with a touch of SDS sometimes.
4. Final wash with >80% EtOH only.
5. Allow beads to nearly completely dry.
6. Elute, with hydrophilic buffer (with or without heating).

Hope that helps
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