Magnetic Bead-Based RNA Extraction

Thumb default avatar
Dec 01, 2016
0
1

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

1 Comments

Thumb default avatar
drshabba 10 months ago

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

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