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
, t2 Y2 I8 H1 h- y  |Baylor College of Medicine, Houston, Texas
/ D2 G# O5 W% `) j# w. t0 U) c! B, B) ?9 X# R5 `1 l
Picking ES Cell Colonies and0 i+ e! b/ L" q1 m
Transferring Them to 24-Well Culture Dishes) O- j4 @: ~  g5 I

/ q  z! e! f% ~2 a& j/ g: F9 g/ y  L/ \# E1 k; m2 P' W
        1.        Spot 15 - 25 ul of trypsin in a 24-well array onto the lid , }) C* p( ?7 M7 d+ J7 H
of a 10 cm plate.1 P' r+ J, @3 }4 T8 k

% c2 t/ m% E6 E) h7 _5 W        2.        After flooding the plate with PBS, pick the colony from % p4 q! G& ]( G2 A( R: b
the plate using a Pipetman set at 2 ul, and transfer it into the trypsin.  
7 g4 p' t' V3 z  `( g0 _" w: |Leave in trypsin for 10-15 minutes.
! {2 _  N4 N+ u# @+ i) `$ P* A$ I5 W, K9 U" }
        3.        Add 20 ul of media from the refed feeder plate to the 4 w9 F; s8 E: d2 w( C8 j6 W
trypsin spot.  Pipet up and down to break up the colony.# z/ k, L5 a8 ^/ p6 w  p5 y

+ @2 F9 U6 G' H. O; P2 f        4.        Transfer the ES cell suspension (approximately 37 ul) # i* A- y$ n! R% Z
into the well of a 24-well feeder plate.
+ [3 j! t3 z; L( P! a
2 Y% H/ c4 Z5 X# w1 g! {& p        5.        Alternatively, 50% of the cells may be lysed directly
; e$ D) C; a: [. L7 @8 J% w0 ufor PCR at this point.  Add 15 ul of the cell suspension directly ( D8 W6 G6 ]- o: @9 a( H" a- j6 K
to 25 ul of PCR Lysis Buffer.
/ l4 H0 \% o. k1 c; _6 j, w
- s- a# x' |4 b- @" `  L4 u* X! n8 _1 C! {& b
From the Laboratory of Dr. Allan Bradley                      8 i7 v. O' }/ M
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