Song Chen:Daily/NOTES/2014-3-6: Difference between revisions

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<b>send H9 in NHSM</b>:
<b>Send H9 in NHSM</b>:
:1*T25 flask to UCLA via Fedex(800555872882)
:1*T25 flask to UCLA via Fedex(800555872882)



Revision as of 22:40, 7 March 2014

hES change medium:

  1. PGP iPS Brainbow knockin #1, #2-4 and #2-6 p2, 2 well*6 well plate in hES/NHSM, respectively;
  2. H9 p62 2*6 cm+24 well*48-well plate in NHSM
  3. H9 Bb6 2 well*6 well plate
  4. PGPs iPS LCB+CRE p62 1*6 cm in hES medium


Send H9 in NHSM:

1*T25 flask to UCLA via Fedex(800555872882)


DE differentition(H9 p62, for methylation analysis-Dinh):

3 well*6-well plate, hES at d2, ActA100 d4


check default GFP expression in knockin iPS lines: Setting: Expose 1209 ms, Cerc, 20x obj,GFP/Brightfield, KI(knockin)

KI#1 KI#2-4 KI#2-6 Bb#6(random integrated)
File:030614 KI-1 GFP.png File:030614 KI-2-4 GFP.png File:030614 KI-2-6 GFP.png File:030614 Bb6 GFP.png
File:030614 KI-1 BF.png File:030614 KI-2-4 BF.png File:030614 KI-2-6 BF.png File:030614 Bb6 BF.png
File:030614 KI-1 Merge.png File:030614 KI-2-4 Merge.png File:030614 KI-2-6 Merge.png File:030614 Bb6 Merge.png

Knockin iPS cell lines don't express default GFP in every cells as expected. #2-4 has no detectable GFP expression, while #1 and #2-6 have a few GFP positive cells. And when compared with brainbow random integrated cell line #6, GFP is expressed at a relative low level, which probably due to the nature of single copy insertion of knockin. The possibility of fluctuation in brainbow expression between knockin cells or contamination with non-transgenic iPS cells await further verification.