Saturday 07/05/14

Posted on July 5, 2014 by admin

10:30 am – 9:00 pm

STORM imaging today

Sequential hybes

  • repeat staining of F06 not much improved on BXC-F06 try 2. Hardly see any stain (just background?) Attempting to image these anyway.

Imaging new stain: D08toD12

  • stains look great. Switching looks great.

Reading

  • interesting article on twist from M. Baylies group
  • a good page on smFISH FAQ from the Raj lab

Project 2

  • start time 1:45 pm.
  • re-clean and paper RNase free zone upstairs
  • making probes.
    • E1 8 wells A01 + A02 — combine and use 25 ug columns
    • E5 8 wells A09 + A10
    • E25 1 well E01 + E02
    • E26 1 well E03 + E04
    • E27 1 well E05 + E06
    • E28 1 well E07 + E08
    • E29 1 well E09 + E10
    • E30 1 well E11 + E12
  • Make dilution of primer plates.
    • Stock concentration = 200 uM
    • need 1:40 dilution. Do 4 in 156.
  • Master mix
    • 25 uL Hot-start Phusion
    • 5 uL 1:100 library
    • 5 uL 5 uM
  • In parallel, dilute my primers 4 in 78 (10 uM working concentration)

Oligo secondaries

  • working on methods description for simulations.

Data analysis

  • picking out regions of yellow chromatin to illustrate scaling
  • launched spot fitting data on recent BXC attempts.
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