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RNA precipitate in black color


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#1 hianghao

hianghao

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Posted 23 February 2011 - 06:30 AM

I've precipitated my RNA in 2 V abs EtOH and 0.1 V sodium acetate 3M pH5.2 o/n at -20C. After centrifugation, black pellet was formed! I thought it was suppose to be white? Is that normal? Also, after washing and resuspend in RNase-free H2O, the purity decrease from 1.x to 0.5...... Please check my protocol:

Add 1/10 volume of Sodium Acetate (3 M, pH 5.2).
Add 2 X volume 95% ethanol.
Incubate o/n -20C
Centrifuge at 14kg 30 minutes at rt
Decant supernatant
Wash with 70% Ethanol twice
Discard supernatant, air-dried 15m
Dissolve pellet in water.




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