Material

  • Medium
  • Deep well plates
  • Liquidator tips
  • Adhesive aluminium seals
  • Adhesive breathable seals

Protocol

1. Sample prep
  • Thaw the 96-well glycerol stocks on the bench wrapped with Kim-towel.
DHFR12-96G010
DHFR12-96G011
DHFR12-96G012
DHFR12-96G013

DHFR3-96G004
DHFR3-96G005
DHFR3-96G006
DHFR3-96G010

*Once thawed, spin samples at 1000 x g for 1 min.

  • Remove seal.
2. Medium prep (Can do while waiting for sample thaws)
  • Take out medium to bench.
  • Label plates based on samples used.
DHFR12-96G010
DHFR12-96G011
DHFR12-96G012
DHFR12-96G013

DHFR3-96G004
DHFR3-96G005
DHFR3-96G006
DHFR3-96G010

*With a liquidator, aliquiot 1000µL of medium to each well (200 µL x 5).

3. Inoculation
  • Change volume of liquidaotr to 10 µL.
  • Take corresponding sample and deepwell plate filled with medium on liquidator.
  • Put on tips, go to glycerol stock.
  • Pippette up and down 2-3 times, and take 10 µL of sample.
  • Move to deepwell plate, and add the 10 µl of glycerol stock to the plate.
  • *Pippette up and down 2-3 times.
  • Discard tip.
  • Culture sample at 37˚C, 160 rpm (circular rotation) for 2 O/N.

Dan Yamamoto-Evans 2019/11/30 16:11

  • interactome/bac96culture.1575098372.txt.gz
  • Last modified: 2019/11/30 16:19
  • by dan