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 # A. L- e' {6 J2 J
Baylor College of Medicine, Houston, Texas
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% U  ]: }$ m" E' t' y% f! `Picking ES Cell Colonies and
! R: n" ^. s7 ~; X+ TTransferring 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
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        2.        After flooding the plate with PBS, pick the colony from % }; o6 @) q) j4 f& g; x
the plate using a Pipetman set at 2 ul, and transfer it into the trypsin.  ) m* H5 a6 ?& Y2 o6 B0 [
Leave in trypsin for 10-15 minutes.' Z8 P0 \% Y0 z* Q$ \1 i& R

! s# H% W. @/ H! G/ F9 ]        3.        Add 20 ul of media from the refed feeder plate to the
* ^. r: v. w3 ctrypsin spot.  Pipet up and down to break up the colony.5 g! J6 g; c, ~' h, y
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        4.        Transfer the ES cell suspension (approximately 37 ul) : S7 s/ G! @2 c8 U4 m6 I5 n
into the well of a 24-well feeder plate.2 L' x0 m  ?% r  z9 j4 I, H

8 v! R* \" j5 d        5.        Alternatively, 50% of the cells may be lysed directly 9 m3 o% x3 e8 {
for PCR at this point.  Add 15 ul of the cell suspension directly
8 ?0 V- i; D9 l1 pto 25 ul of PCR Lysis Buffer.
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; Q/ W8 Z/ d% y; z) KFrom the Laboratory of Dr. Allan Bradley                      5 s4 K7 w3 A, \: Y" t
Baylor College of Medicine, Houston, Texas

sacia

学习了，谢谢楼主啦

llingzu

kankan

可怜的孩子

谢谢啊