Monthly Archives: May 2013

10:00 A – 7:00P, 8:30P – 12:40A File maintenance Alistair12 out of disk space! Black-DNA region run crashed due to running out of disk space STORM analysis crashed due to no free disk space to save files STORM analysis Fab7 … Continue reading →

8:00 PM – 11:20 P probe making Set up new test gel: 3R4500, Ubx, AdbdA, AbdB, BX-C run OPAINT, and OPAINT1 72C cell staining plate new Kc167 cells on coverslips Fix run through all prep steps up to 50% formamide … Continue reading →

10:10 A – 5:30 P RNA labeling of cultured cells incubate cells in 55C RNA hybe buffer, 30 min transfer to RT 2x SSC, wash 10 min transfer to PBT wash 10 min+ 2x block 1 hour stain with shp … Continue reading →

11:00 A – 10:00 P Probe and hybe troubleshooting check staining AATAT control and Y look good black maybe stained, check after washout low and high RNA-en-P1 both have substantial nuclear background, but no clear signal Coding Fixed bugs in … Continue reading →

11:00 A – 7:30 P, 9:00 P – 11:00 P Trouble-shooting hybes check samples: AATAT-P1 + S1-short on cells – strong and clear en-P1 + S1-short on cells – clear staining, bleaches fast en-RNA-P1rc + S1-short on cells – no … Continue reading →

10:20 A -7:30 P Project 2 Piranose: staining is still good in piranose treated cells. GA background is EXTREME, but certainly uniform, not punctate on the whole cell conventional level at least Cell staining check cells from yesterday — all … Continue reading →

10:30 A – 8:35 P Probe making resuspend ethanol precipitated probe samples make and cast new UREA gels. 2x 15 well .75mm, 1x 5 well 1.5 mm (25 mL is not enough for 2x 1.5mm — these take 10mL each. … Continue reading →

11:10 A – 7:30 P, 9:30P – 10:30 P probe making Labeling amine-tagged probe with cy5 dye. labeling reaction for secondary cy5 (see protocol) labeled precipitated DNA pellet awaiting HPLC purification (need directions from Hao). Test large scale reactions with … Continue reading →

10:20 A – 7:15 P, 8:30 P – 1:45 A cell staining remove samples from 40C, place in RT 2x SSCT for first rinse out. check staining of control slide on STORM2 TIRF- no staining except in dead cells (should … Continue reading →