LAMBDA DNA MAXI-PREP

pre-grow BNN 45 in LB plus .2% maltose
centrifuge and concentrate in SM
measure OD600 and calculate cells/ml (assume OD= 1 is 8 x 108)
in phage tube add: 1 x 1010 cells
5 x 107 phage
adjust volume to 3 mls with SM
incubate 37 C x 20'
innoculate pre-warmed NZ liquid media. (1 L)
incubate 37 C for 9-12 hours, lysis should be evident. If not, quit!
add 10 mls CHCl3, incubate 37 C x 10'
centrifuge 5 kg x 30', decant supernatant and cool to room temp.
add DNase and RNase, both to 1 ug/ml.
room temp. x 30'
add NaCl (solid) to 1M. (58.4 g/L)
dissolve, then ice x 1 hr
add PEG 6000 (solid) to 10% w:v (100 g/L)
ice x overnight
cent 5 kg x 30', discard supernatant
resuspent pellet in 2 ml SM
extract once with CHCl3
adjust volume to 4 mls with SM, add 3.0 g CsCl
split into 2 TL 100 tubes
cent. 100 krpm x >6 hrs, 20 C

NOTES:
1. isolate phage band, remove CsCl, and isolate DNA as described in
Maniatas.