Sera-mag buffers from Illumina RNA-seq kit

[brooksma]

Does anyone know the recipes for the Sera-mag oligo dT Bead Binding and Wash Buffers from the Illumina mRNA-seq Kit? I'd like to use the extra beads fro the kit to purify mRNA to optimize library preps and would like to keep the mRNA purification step the same. Thanks.

[Paulo.jt]

Hi, I´m looking for these buffers as well. Does anybody know the recipe?

[brooksma]

Hi Paulo,
After talking to Illumina tech support (they were pretty tight lipped about the recipe) I was able to get at least the components but not the concentrations. I looked around at different protocols and came up with my own recipe. I've tried it and it seems to work just fine. My frag profiles from BA2100 look identical when done on the same RNA sample using Illumina's Buffers. Here is what I made:
Binding Buffer (200mM Tris, pH7.5, 1M LiCl, 2mM EDTA, 0.2% Tween-20), Wash Buffer (100mM Tris, pH7.5, 150mM LiCl, 1mM EDTA, 0.1% Tween-20).
Good luck,
MB

[mapplebe]

Are you sure that you used 200 mM Tris? I have been looking at several different buffer recipes posted around the internet, and they all seem to use 20 mM Tris / 10 mM Tris in the washing/binding buffers but are otherwise identical (except that the tween may be optional)

[protist]

Originally Posted by brooksma

Hi Paulo,
Here is what I made:
Binding Buffer (200mM Tris, pH7.5, 1M LiCl, 2mM EDTA, 0.2% Tween-20), Wash Buffer (100mM Tris, pH7.5, 150mM LiCl, 1mM EDTA, 0.1% Tween-20).
Good luck,
MB

Aside from the Tris and tween the recipes are exactly the same as those used for Dynal oligo(dT) beads used for polyA selection:
Binding: 20 mM Tris-Cl pH 7.5: 1 M LiCl: 2 mM EDTA
Wash: 10 mM Tris-Cl pH 7.5: 0.15 M LiCl: 1 mM EDTA