CsCl-EtBr plasmid prep.

GWDGV1::KCHOWDH kchowdh at gwdgv1.dnet.gwdg.de
Wed Dec 1 12:07:20 EST 1993


Hi, Here is the method that always works for me.
Remove the band witha syringe
Extract 2-3 times with water saturated butanol
dilute the EtBr free aqeous phase 3 fold with TE to avoid the ppt.of CsCl in
a corex tube
add NaCl to 0.1M or NaAc to 0.3M, mix
add equal vol. of Isopropanol or 2.5 vol,. of EtOH,mix
put at 4 C or -20C for 1h- overnight depending on the amount of DNA
Spin at 10000rpm in Sorval for 15 min
The DNA is in the pellet
Wash pellet with 70% Etoh, dry and suspend in TE
This DNA might also contain nucleases, So it is better to extract with PCI
and reppt.
Hope this helps.
Cheers,
Kamal.



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