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Concentrating poly(A)+ mRNA

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Concentrating poly(A)+ mRNA

 
Source: Protocol Online
Abstract: Simple precipitation method of concentrating poly(A)+ mRNA.

Procedure

  1. Add 1/10 volume 3 M NaAc, pH 5.2, and 2.5 volumes 100% 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.

 

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