Nucleic acid concentration and/or cleaning

Protocol used to concentrate poly(A)+ mRNA, RNA, DNA. Also to clean reactions from buffers and others that may interfere in dowstream reactions.


1. Add 1/10 volume 3 M NaAc (Sodium acetate) and 3 volumes of cool ethanol 95% (high purity bottle).

2. Mix well and incubate at -20°C for at least 20 min.

3. Centrifuge at > 12,000 x g in a microfuge for 20 minutes (if RNA work at 4°C.)

4. Discard the supernatant and wash the pellet twice with 70% ethanol (500 uL, 3 min centrifuge).

5. Air dry pellet. Check for dryness before proceeding. (no ethanol should be seen)

6. Resuspend pellet in RNase-, DNase-free H2O. Shake well, spin 5 seconds a couple of times. Let stand in ice 20 minutes.

The appropriate volume for resuspension depends on the expected yield and the amount of RNA/DNA required for your experiment (like for electroporation dissolve in 5 uL, for miniprep 30-50uL).

7. Store the solution at -80°C/RNA, -20°C/DNA.

8. If needed, quantitate by spectrophotometry and electrophoresis.

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