96-well E.coli culture
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