Template:SBB-Protocols Assay1: Difference between revisions

From OpenWetWare
Jump to navigationJump to search
No edit summary
Line 29: Line 29:
===='''Monday'''====
===='''Monday'''====
*Run the assay, starting from the 384 well plate step.
*Run the assay, starting from the 384 well plate step.
*create
*Plate all the constructs and the controls.


===='''Tuesday'''====
===='''Tuesday'''====
*Pick 10 colonies from each plate and grow on the 96 well block to saturation.
===='''Wednesday'''====
*Run the assay from the 384 well plate step.

Revision as of 12:02, 15 April 2009

Growth Assay Protocol

  • Replate 16 constructs on new plates at the same time.
  • Plate DH10B, pBca9145-Bca9494, and pBca9495CA-Bca1144 at the same time.
  • Grow plates overnight.
  • Pick 5 colonies from each sample.
  • Grow to saturation in 96 well blocks with 400 uL LB media in each well.
  • Then from each of the 5 unique liquid cultures, make an arabinose sample and a non-arabinose sample.
  • Add 50 uL of LB media per well in 384 well plate.
  • Add 1 uL of cell sample to each well.
  • Add 1000x Arabinose to 5 out of the 10 wells for each construct.
  • Place plate in Tecan and run OD measurements every 10 minutes.

Schedule

  • For the first run-through, we'll do the 3 controls.
  • Get one of the amazing GSI's to pick 10 colonies of each control Sunday night, suspending each colony in 400 uL LB media in 96 well plate.

Monday

  • Run the assay, starting from the 384 well plate step.
  • Plate all the constructs and the controls.

Tuesday

  • Pick 10 colonies from each plate and grow on the 96 well block to saturation.

Wednesday

  • Run the assay from the 384 well plate step.