Lab 2.1 prep
Lab 2.1 prep kdorfman Wed, 08/25/2021 - 20:449/16 & 9/21 2021 9/20 & 22 2022
- H2B-mCherry (Addgene 20972) (Amp resistant) (Labeled A)
- ecadherinGFP (Addgene 28009) (Amp resistant) (Labeled B) GROWS SLOWLY! ALLOW EXTRA TIME!
| for Th lab | for Tu lab | prep |
|---|---|---|
| Monday 9/13 | same | streak each strain onto LB Amp plate; incubate ON |
| Tues 9/14 | same | plates to fridge; re-streak if necessary |
| Wed 9/15 | Mon 9/20 | pick colony, start liquid ON culture in 100 mL LB-Amp (one of each strain) |
| Th 9/16 | Tu 9/21 | Concentrate cells (max speed >10 min), then aliquot ~1 mL per pair of each strain |
Miniprep 2022
Tuesday: Zyppy (use up old aliquots)
Thursday: ZR Classic
Mini-Prep 2021
- micro pipetters & tips
- labeled racks in fridge
- 1 mL each E. coli culture (concentrated by centrifugation) per pair
- centrifuges
Miniprep kits - aliquot miniprep reagents per pair:
- sterile microfuge tubes
- 250 uL Buffer P1(+ RNaseA+LyseBlue)/rxn = 510 uL per pair
- 250 uL Buffer P2/rxn = 510 uL per pair
- 350 uL Buffer N3/rxn = 710 uL per pair
- 2 QIAprep spin columns
- 0.5 mL Buffer PB/rxn = 1.1 mL/pair
- 0.75 mL Buffer PE/rxn = 1.51 mL
- 50 uL Buffer EB (10 mM Tris-Cl pH 8.5) per rxn = 110 uL per pair
- sterile water