Isopropanol precipitation of DNA.

Nigel Kenward mbxnjk at vax.nott.ac.uk
Thu Oct 26 07:31:27 EST 1995


Hi,
   I routinely use 1 volume of isopropyl alcohol (IPA) for 
precipitation....I add 0.1 volumes of 3M NaOAc, 100mM Mg2(OAc)2, pH5.2 to 
aid precipitation. If I want to sequence the DNA I usually reprecipitate 
with an equal volume of 800mM NaCl, 13% PEG 800. Incubate on ice for 30 
mins to precipitate DNA, spin for 10 mins at max. in a microfuge, wash 
pellet (care!! It's translucent and comes away from the tube easily) with 
70% ethanol, dry and re-dissolve in TE. Gives excellent DNA and it's 
recommended by ABI for use in sequencing reactions with their automated 
sequencers.
  Have fun!


              Simon.

-- 

Dr. Simon Dawson           TEL:+44 (0)115 9249924 ex. 44789
Dept. of Biochemistry      FAX:+44 (0)115 9422225
Queens Medical Centre      Email:Simon.Dawson at .nott.ac.uk
Clifton Boulevard          URL: http://www.nott.ac.uk/~mbxspd/Simon.html
Nottingham
U.K.                       "Back off man, I'm a scientist!" - Bill Murray.



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