TY - JOUR
T1 - Preparation of DNA from embryonic stem cells or other cultured cells
AU - Behringer, Richard
AU - Gertsenstein, Marina
AU - Nagy, Kristina Vintersten
AU - Nagy, Andras
N1 - Publisher Copyright:
© 2017 Cold Spring Harbor Laboratory Press.
PY - 2017/12
Y1 - 2017/12
N2 - Characterization of embryonic stem (ES) cell-mediated genome alterations, including random insertional transgenesis, gene trapping, gene targeting, and site-specific recombinase-mediated changes, is performed mostly at the ES cell level, before the introduction of these alterations into a mouse. A detailed characterization requires a larger amount of DNA than is required for the initial detection of the candidates for the desired alteration. This protocol describes the preparation of DNA from a 10-cm tissue culture plate. The cells are trypsinized and lysed, and DNA is recovered from the lysate by organic extraction followed by ethanol precipitation.
AB - Characterization of embryonic stem (ES) cell-mediated genome alterations, including random insertional transgenesis, gene trapping, gene targeting, and site-specific recombinase-mediated changes, is performed mostly at the ES cell level, before the introduction of these alterations into a mouse. A detailed characterization requires a larger amount of DNA than is required for the initial detection of the candidates for the desired alteration. This protocol describes the preparation of DNA from a 10-cm tissue culture plate. The cells are trypsinized and lysed, and DNA is recovered from the lysate by organic extraction followed by ethanol precipitation.
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U2 - 10.1101/pdb.prot092718
DO - 10.1101/pdb.prot092718
M3 - Article
C2 - 29196598
AN - SCOPUS:85037053509
SN - 1940-3402
VL - 2017
SP - 1016
EP - 1018
JO - Cold Spring Harbor protocols
JF - Cold Spring Harbor protocols
IS - 12
ER -