Liquid culturing protocol for agar art E. coli in 5mL volumes

	For N=4: (At BosLab: Frank Lee and Ambar Sarkar on 17-July-2021, and Frank Lee on 20-July-2021)
2 × N sterile culture tubes with caps	10 culture tubes = 8 culture tubes + 2 control tubes FYI 16-July-2021: Boslab has plenty of 15 mL culture tubes.
2 sterile culture tubes, with caps, as controls: 1 LB only 1 LB + chloramphenicol only
Rack/holder for above culture tubes	We used 2 racks with 12 open spaces each so I could start with all 10 tubes on one rack. Then as I perform a protocol step on a tube, I can move it to the other rack to indicate that that tube has completed the current step. This helps me track my progress in case I get distracted while performing the protocol (e.g. a lab partner needs help).
Marker pen for labeling tubes	Use a VWR Lab Marker
Stock bottle of at least (N+1) x 10 mL of sterile liquid LB media	Liquid LB amount needed: 60 mL = 50 mL plus 10 mL extra just in case
1 sterile serological pipette to aliquot (N+1) x 10 mL	will use a 50 mL sterile disposable pipette
1 sterile serological pipette to transfer 5 mL	will use a 5 mL sterile disposable pipette
pipettor for serological pipettes (e.g. pipette-aid)	will need to check that the pipettor doesn't leak and has good batteries
1000X chloramphenicol stock solution, 25 mg/mL, from the −20°C freezer (they are still liquid at −20°C)	1000x chloroamphenicol stock solution needed: 60 µL (i.e. use 1 µL stock solution per mL of media)
micropipettor to deliver N × 10 µL	use a P200 (i.e. 200 µL capacity pipettor)
1 sterile micropipette tip for the above	get the box of P200 tips (I realize they've been opened and aren't guaranteed to be sterile – I think the chance of contamination from a new P200 tip is small – though if the protocol fails we'll try pipetting 70% ethanol first before retrying.)
sterile culture media bottle to hold the LB media aliquot mixed with chloramphenicol	an empty, clean, autoclaved 100 mL glass media bottle
(2 × N + 2) sterile inoculation loops all from the same bag or container	10 sterile inoculation loops
Plates with streaked, colorful, agar art colonies from the fridge or incubator	the 4 single-color agar plates from the fridge marked “Frank Lee”
Eppendorf tube of 70% ethanol for sterilizing a P200 tip.	Use a 5 or 10 mL serological pipette to aliquot an inch or so of 70% ethanol into a 1.5 mL Eppendorf tube. Use this as an opportunity to test that your serological pipettor has good batteries and doesn't leak.
Laminar flow hood	reserve the Boslab PCR Workstation

¶ Revised 2021 July 20 by Frank Lee