Tag Archives: confocal

Monday 06/05/13

9:30 A – 7:45 P, 8:45P – 11:55 P Oligopaints Running out gradient gels (one at a time) hotter is better with mis-priming. using 20 bp PPT primer and library specific NP primer: with adapters: Check Ph stains booked confocal … Continue reading

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Sunday 01/13/13

3:30P – 11:55 P Fly work Flipped most of rescue crosses.   Several vials extinct.  Should still have more vials than I want to screen though. New crosses of SCM males to MTD virgins (2 vials). Examine current crosses of … Continue reading

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Friday 01/10/13

8:30 A – 9:50 P Fly work Flip collection plate 8:30A Fixing embryos 11:30A.  (20 min 8% FA reaction) Flip all polycomb mutants into bottles STORM made new bead slide, 1:25K imaging 2 rounds of beads for 3D fitting K9me2 … Continue reading

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Wednesday 01/09/13

10:00A — 10:45P Cell culture blocking IF first.   secondaries for IF first (2 m-750 (6:1), 6 wells m-488 (2.0)) 10 min in .1% GA, 4% PFA post fix.  wash in PBT, then 2xSSCT following Brian’s protocol.  Need rubber cement. … Continue reading

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Tuesday 12/11/12

10:10 A Check PCRs (in blocks 3A (sim screening) and 3B (Espl screening) better way to do this: complementation test back to original sim[D] and Espl[D] lines.  Just need to expand those lines back to bottles, will need ~100 vials … Continue reading

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Saturday 12/08/12

11:00 A – 7:30P, 8:45P – 10:00P Cell staining label with m-HP1a -405,750 and rb-H3K27me3 -488 (top) or rb-H3K27ac -488 (bottom row) Polytene cell staining label with m-HP1a-Cy3b and rb H3K4me3 -488 or rb H3K27me3-488. check staining on confocal some … Continue reading

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Thursday 12/06/12

9:45 A – 7:30 – 11:55P Troubleshooting antibodies with polytene H3K9me2 failed completely on polytene cells.  Dm01 control labeled in parallel looks bright and clean (better than direct labeled Dm01). Polytene labeling today: testing H3K27me3 (see if polytene chromosomes label … Continue reading

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Wednesday 12/05/12

9:30 A  – 7:40P, 9:00P – 12:00A Confocal dot distances Finish O/N confocal run of Ubx-AbdA data Probe making sent order to LC Sciences for chromatin types On why previous probe might have failed:  over drying? — DNA and RNA … Continue reading

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Tuesday 12/04/12

9:40 A – 7:45P, 9:00P – 1:00A Dfd/tub analysis Break down O/N confocal run 3 channels, speed 6, 22 positions, 17.5 finished in 11 hour run.  contact zeiss rep Casey about changing channels between z-stacks rather than every section, should … Continue reading

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Monday 12/03/12

8:45 A — 7:30 P, 8:45P – 12:40 A Confocal data Finish O/N confocal run transfer files modify imviewer LSM to automatically detect all .lsm files in a folder and convert them all to TIF / labeled stacks of TIFs … Continue reading

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