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Summary: UCSD Transgenic Mouse and
Gene Targeting Core
9500 Gilman Drive, MC 0687
La Jolla, CA 92093-0687
(858) 534-3178
Current as of 3/01 1
DNA Isolation from ES cells in 96 well plates
After the clones from your transfection have been picked and split, you will receive
duplicate 96-well plates that have been grown to 100% confluence. The media will have
been removed and the plates will have been frozen at 20o
C. You should prepare DNA
from one of the plates first and store the duplicate covered with parafilm at -20o
C as a
backup.
Required Reagents and Materials:
1. Lysis Buffer For 100 ml:
10 mM Tris-HCl pH 7.5 0.5 ml of 2 M stock
10 mM EDTA: 2 ml of 0.5 M stock
10 mM NaCl: 0.2 ml of 5 M stock
0.5% Sarcosyl w/v: 0.5 g / 100 ml
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