9:30a – 8:30p
Banff to do
- email speakers who haven’t sent in titles (individual emails sent).
- update schedule on website with tentative speaker order
- both goals complete for today
Probe making
- oligo cleanup of probes
- new gel from O/N gel-green looks much better:
- by nanodrop no substantial increase in yield from 10ug to 20ug primer. It does look like all the primer got used in 20ug AbdA but not the 10ug (will be better with labeled primer).
- probes are 119 bp long (53 adapter + 42 probe + 21 primer + 3 T7 Gs).
- at an ave. of 500 ng/uL, but this is a mix of primer and probe. If it was all probe it would be ~12 pmol/uL
- had 300 pmol of primer. Recovering all of this would have given 350 ng/uL
- 150 ng/uL of 66 more bases = 7 pmol/uL
Cell culture
- KC-cells not attaching! Need to baby cells back up to happy / healthy?
- use last 2 coverslips from previous batch of Kc-167 cells to test new Ubx probe (with RNase). Use 2nd coverslip for no-primary control.
- prepping discarded Pc and Psc stained S2 cells for DNA-FISH (5 coverslips)
- start last vial of Kc167 cells from stock. Be sure to make new stock of these. Be sure to passage every 4 days or less.
- new stock is adhering properly during recovery. Let’s hope it continues to behave well.
- Try a complete media change with existing Kc167 cells.
Cell staining
- ubx-20 5uL + 1uL RNase + 1.2 uL secondary dock + 0.6 uL 405 + 20 uL hybe buffer.
- control = no primary, 1uL RNase + 1.2 uL secondary dock + 0.6 uL 405 + 20 uL hybe buffer.
other
- helping Hao get set up wit Github and Matlab STORM