Haynes Lab:Notebook/RNA-seq of PcTF Transfected U2OS & SK-N-SH cell lines/2015/01/19

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Since the U2OS, SKNSH and KAH126/U2OS cultures were expanded/passaged on Friday last week, today I'll be checking on the cultures to observe their progress. The KAH126/U2OS culture in the 6-well plate should be approaching 80% confluency, at which point I'll replace the medium with fresh medium and induce with doxycycline.


  • Check U2OS, SKNSH, and KAH126/U2OS cultures for confluency
    • If KAH126/U2OS culture in 6-well plate is 80% confluent, induce expression w/ doxycycline


To induce KAH126/U2OS cells: add 1μL doxycycline from the freezer per mL of culture. Mix by gently shaking, then place culture back in the incubator.


Observations, measurements, placeholder info etc. go here


SKNSH culture (PS-β) - at approximately 20% confluence. Needs a few more days.

U2OS culture (PS-β) - over 80% confluence. Passage into PS-1 and split off a 6-well plate tomorrow.

KAH126/U2OS culture (6-well plate) - 100% confluence. Missed the deadline for inducing doxycycline expression. Need to make another 6-well plate from the stock, meanwhile I'll be inducing this plate to see what happens. I'm inducing 4 wells - 2 for day 5 sampling, 2 for day 10 sampling, leaving 2 for negative controls.

KAH126/U2OS culture (PS-2) - over 90% confluence. Tomorrow I'll passage to PS-3 and split some off for another 6-well plate. This time do a regular dilution - 0.2mL of parent culture per 4mL of new well.