Picking ES Cell Colonies and Transferring Them to 24-Well Culture Dishes

肖爱华

Picking ES Cell Colonies and Transferring Them to 24-Well Culture Dishes From the Laboratory of Dr. Allan Bradley 6 q5 ^+ L0 [9 L6 O$ p
Baylor College of Medicine, Houston, Texas3 a1 D3 ]5 T8 L5 I& w8 w, F

4 X* Y4 s' c+ J3 t- e& yPicking ES Cell Colonies and
% u. Q$ c' j5 y- zTransferring Them to 24-Well Culture Dishes
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        1.        Spot 15 - 25 ul of trypsin in a 24-well array onto the lid 2 |* V( N; u+ L0 p" P
of a 10 cm plate.1 ^8 Q( D4 B! }4 r" }" g$ B
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        2.        After flooding the plate with PBS, pick the colony from
7 {5 Q( n0 b' l2 @3 {( mthe plate using a Pipetman set at 2 ul, and transfer it into the trypsin.  
: K) S4 O% n" j- ]2 `) W) w6 _Leave in trypsin for 10-15 minutes.
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5 s6 B5 N$ V2 O, T+ [. F        3.        Add 20 ul of media from the refed feeder plate to the . ^$ D. C. C, T$ Y6 G; a
trypsin spot.  Pipet up and down to break up the colony.) T; q/ ~7 s7 t2 q1 i
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        4.        Transfer the ES cell suspension (approximately 37 ul)   y/ n  b4 u) g: G
into the well of a 24-well feeder plate.
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" c, F8 J$ Y' Q& Q% N/ @$ C' N        5.        Alternatively, 50% of the cells may be lysed directly 8 O1 h* f9 F. a) v4 P3 \
for PCR at this point.  Add 15 ul of the cell suspension directly 1 ?( F  O( }# o
to 25 ul of PCR Lysis Buffer.
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6 d/ D% q: A6 j" |% C* k7 }2 j/ I+ P+ S( gFrom the Laboratory of Dr. Allan Bradley                     
$ [# Y/ P% x2 @# p/ R1 ~Baylor College of Medicine, Houston, Texas

可怜的孩子

谢谢啊

llingzu

kankan

sacia

学习了，谢谢楼主啦