How to concentrate poly(A)+ mRNA - how to cocentrate mRNA (Jun/27/2004 )

how to cocentrate mRNA thanks

Hi.

you should firstly precipitate mRNA with 1/10 th volume 3M Sodium Acetate, 1/50 volume Glycogen(5mg/ml), and 3 volume ice cold 100% ethanol at -20C for at least 1hr, then resuspend the precipitate in TE buffer(pH 8.0) .

Good luck

weimin

Here is a protocol used to concentrate poly(A)+ mRNA:

1. Add 1/10 volume 3 M NaAc, pH 5.2, and 2.5 and 2.5 volumes ethanol.
2. Mix and incubate at -20C for at least 1 hour.
3. Centrifuge at > 12,000 x g in a microfuge for 20 minutes at 4C.
4. Wash the pellet twice with 80% ethanol.
5. Air dry pellet. Check for dryness before proceeding.
6. Resuspend pellet in RNase-, DNase-free H2O. The appropriate volume for resuspension depends on the expected yield and the amount of RNA required for the cDNA synthesis.

Reference: Affymetrix Expression Analysis Technical Manual.