Plasmid Q membrance Purification 10232012

Plasmid MaxiPrep Q Membrane using Mustang Q menbrance.

10/23/2012

Using protocol: Plasmid MaxiPrep Q Membrane

Sample : 1382, 260ml,  OD=2.04

Adding 10 ml P1 (with RNase) +10ml P2+10ml P3, following the protocol except washing buffer B (0.75 M NaCl).

Elute with buffer C (1.6M NaCl), collect 1ml fraction 1, 5 ml fraction 2 and 4 ml fraction 3. measure DNA using nanodrop.

Fraction 1= 3ug/ml: fraction 2= 48.7ug/ml, fraction 3=10 ug/ml

combine fraction 2 and 3 (280 ug DNA)  and precipitate with 0.7 Volume isopropanal.

Centrifuge at 15000 rpm for 25 mins.

Can see a  little white, glassy pellet, wash with 70% ethenal, dissove in 300 ul TE buffer. The conc. is 100ng/ul.  totally get 30ug DNA, DNA recovery is very low, about 10-15 % after precipitation.

need to dissole in less volume to get high Conc. for next step transfection.