Song Chen:Daily/NOTES/2014-3-6: Difference between revisions
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>Chensong (Created page with "<b>hES change medium</b>: #PGP iPS Brainbow knockin #1, #2-4 and #2-6 p2, 2 well*6 well plate in hES/NHSM, respectively; #H9 p62 2*6 cm+24 well*48-well plate in NHSM #H9 Bb6 2...") |
>Chensong No edit summary |
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#PGPs iPS LCB+CRE p62 1*6 cm in hES medium | #PGPs iPS LCB+CRE p62 1*6 cm in hES medium | ||
<b> | |||
:1*T25 flask | <b>Send H9 in NHSM</b>: | ||
:1*T25 flask to UCLA via Fedex(800555872882) | |||
<b>DE differentition(H9 p62, for methylation analysis-Dinh)</b>: | <b>DE differentition(H9 p62, for methylation analysis-Dinh)</b>: | ||
:3 well*6-well plate, hES at d2, ActA100 d4 | :3 well*6-well plate, hES at d2, ActA100 d4 | ||
<b>check default GFP expression in knockin iPS lines</b>: | <b>check default GFP expression in knockin iPS lines</b>: | ||
Setting: Expose 1209 ms, Cerc, 20x obj, | Setting: Expose 1209 ms, Cerc, 20x obj,<font color=green>GFP</font>/Brightfield, KI(knockin) | ||
{|{{table}} | {|{{table}} | ||
|[[File: | |KI#1 | ||
|[[File: | |KI#2-4 | ||
|[[File: | |KI#2-6 | ||
| | |Bb#6(random integrated) | ||
|[[File: | |- | ||
|[[File: | |[[File:030614_KI-1_GFP.png|200px]] | ||
|[[File: | |[[File:030614_KI-2-4_GFP.png|200px]] | ||
|[[File:030614_KI-2-6_GFP.png|200px]] | |||
|[[File:030614_Bb6_GFP.png|200px]] | |||
|- | |||
|[[File:030614_KI-1_BF.png|200px]] | |||
|[[File:030614_KI-2-4_BF.png|200px]] | |||
|[[File:030614_KI-2-6_BF.png|200px]] | |||
|[[File:030614_Bb6_BF.png|200px]] | |||
|- | |||
|[[File:030614_KI-1_Merge.png|200px]] | |||
|[[File:030614_KI-2-4_Merge.png|200px]] | |||
|[[File:030614_KI-2-6_Merge.png|200px]] | |||
|[[File:030614_Bb6_Merge.png|200px]] | |||
|} | |} | ||
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 in knockin cells, 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. |
Latest revision as of 22:41, 7 March 2014
hES change medium:
- PGP iPS Brainbow knockin #1, #2-4 and #2-6 p2, 2 well*6 well plate in hES/NHSM, respectively;
- H9 p62 2*6 cm+24 well*48-well plate in NHSM
- H9 Bb6 2 well*6 well plate
- 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)
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 in knockin cells, 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.