9:25 A -5:50 P
Ph FLAG
- Processing confocal images of Ph data
- updated ImageShop / ExtractLSM to fix bug in multi-position loader.
- Ph FLAG discussion / comments: post
Literature
- Pirrotta Insulators target active genes to transcription factories and polycomb-repressed genes to polycomb bodies[http://www.plosgenetics.org/article/info%3Adoi%2F10.1371%2Fjournal.pgen.1003436]
- Bickmore Divergence of Mammalian higher order chromatin structure is associated with developmental Loci
- van Steensel and Bickmore review
- Cavalli and Misteli Review
- Added Bickmore and van Steensel to feedly tracker
- Gro-Seq PolII waves to track elongation rates (Mol Cell)
STORM analysis
- set up en-embryo analysis running on Cajal (10 cores).
Review/Referee
- Submit review to journal office: all previous issues have been thoroughly addressed by the authors.
Probe making
Qiagen Buffers
- Buffer PE
- 10 mM Tris-HCl pH 7.5
- 80% ethanol
- Buffer PB
- 5 M Gu-HCl
- 30% isopropanol
- (My Zymo binding buffer is pH 4.8)
measurements (ng/uL). in 150 uL unless otherwise indicated
- Y2.5 E1 263
- Y2.5 E2 81
- Y1.5 E1&2 272
- K2.5 E1 519
- K2.5 E2 435
- K2 E1 150
- K2 E2 100
- K2 E3 48 (2x + reblank. prev 16?!)
- G E1 97
- G E2 (300 uL) 35
- G2.5 E2 450
- Ubx E1 150 (prev 140)
- Ubx E2 150 (prev 130)
- Ubx E3 48 (prev 16)
- AbdA E1 115
- AbdA E2 120
- AbdA E3 77 (prev 40)
- AbdB E1 115
- AbdB E2 150
- AbdB E3 50 (prev 20)
Nicking reactions (with 1/2x max enzyme concentration)
- 450 uL DNA (AA, AB, Y) + 52uL buffer 2 + 23 uL Nb.BsrD1
- 600 uL reactions (G, Ubx) + 70 uL buffer 2 + 30 uL Nb.BsrD1
- 750 uL reactions (K) + 100 uL buffer + 40 uL Nb.BsrD1
- Incubating 5 hr at 65C in 100 uL aliquots.
- send request to order more Nb.BsrD1