Category Archives: probe and plasmid building

Protected: Alternative library and probe designs

Posted on March 10, 2013 by admin

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Protected: probe design

Posted on February 27, 2013 by admin

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Wednesday 01/16/13

Posted on January 16, 2013 by admin

9:30 A – 7:30 P  Finish comments on counting comparison for ML # Finish revisions Write to MERCK about Probe Making Precipitate and resuspend RNA-FISH DNP probes for tou, inv, en, Antp and Dfd. Make new RNA-Hybe soln to resuspend … Continue reading →

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synthesis of RNA in situ probes

Posted on January 15, 2013 by admin

Set up in vitro transcription reaction as follows: Reagent Volume 5x transcription buffer (ProMega): 4ul 10x hapten (Dig. Bio DNP) U-NTP mix 2ul 100mM DTT 1ul RNase inhibitor (RNasin) (50 μ/μl) 2ul linearized DNA (~250 ng if 0.2μg/μl) 1ul (vary … Continue reading →

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Saturday 01/12/13

Posted on January 12, 2013 by admin

10:30 A – 8:00P Probe making precipitate probe, resuspend in fresh DNAse free ddH2O snail data analysis run nuc mapping cyto2nucmap for MP08Hz_snaD_2 data modify lsm_viewer to not append embryo numbers when running export all on embryos data without position … Continue reading →

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Friday 01/10/13

Posted on January 11, 2013 by admin

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 11/22/12

Posted on November 22, 2012 by admin

9:10 A — 8:00P, 10:30P – 12:45 A UltaCryo collect prepped samples 10A – 2P, cutting ultra cryo sections of 1-10hr MTD DFISH-fixed embryos For next time: denser embryo packing in smaller gelatin sections Cryo protect at least 30 min … Continue reading →

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Monday 11/19/12

Posted on November 19, 2012 by admin

9:15 A — 7:15 P, 8:15P – 1:15 A Cell labeling hot washes for probe first slide (2x at 60C) 1x at RT remove and save primary antibodies at RT for primary first slide. PBT rinses (3x soln change, 3rd … Continue reading →

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Friday 11/16/12

Posted on November 16, 2012 by admin

9:45 A – 7:00P, 8:00P – 10:30P Making summary images of SCM data. Reconcentrating / Precipitating probe prior to nicking reaction. Combined into 2 tubes. Dissolving pellets at 37 O/N.  (waiting for Nicking enzyme to continue). Nicking enzyme arrived Brian … Continue reading →

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Thursday 11/15/12

Posted on November 15, 2012 by admin

9:15 A — 6:45 P, 8:30P – 12:00A collect virgins for sim Espl[D] recombineering new cross, sna males x Sp/CyoZ; TM2/TM6 virgins clear MTDs and TM2/TM6s for virgin collection (peak emergence for latest bottles?) pulverize and refreeze Espl[D] and sim[D] … Continue reading →

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