RNA/DNA/PROTEIN FROM SAME SAMPLE??
andrews at physiol.su.oz.au
Sun May 7 23:47:16 EST 1995
This method is very useful for isolating DNA from large samples and involves the use of
high NaCl. The reference is Miller, SA etal. (1988) Nucleic Acids Research 16 (3)
The protocol is for whole blood, but I am sure that you could adapt it to your needs using
the buffers and reagents normally used for lysis and washing cells.
Basically after the cells have been lysed (in around 3 ml of buffer), 2 ml of saturated
NaCl is added and mixed gently for about 15 seconds. The cellulsr proteins may be pelleted
by centrifugation at 2500 RPM (in an SS34 or JA-17) for 15 mintes at RT. The
supernatant containing the DNA can then be decanted and spun again to remove residual
The protein precipitate may be retained and 2 volumes of ethanol (RT) is added to the DNA
solution. If enough DNA is present it precipitates in front of you and can be spooled to
remove it. If only a small amount is precipitated, it may be spun at high speed in a carex
tube and resuspended in TE or whatever.
I am not aware of a method for isolating RNA from tissue that is non toxic, howver, this
protocol may be adapted in some way for this purpose.
I hope that this helps you out in some way.
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