Team:Paris Bettencourt/Notebook/Riboswitch
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July 27th
Goal
E coli DH5-alpha transformation with RFP from iGEM plate 4 - 2H (pSB1A3 backbone)Procedure
- I took iGEM plate 4 - 2H (RFP, BBa_J04450 in PSB1A3 backbone) and resuspend it in 10 uL of distilled water.
- Used E coli competent cells from NEB 2 tubes c2987 (50uL) : 1 control (C) and one for transformation (T)
- Introduced 2 uL (200-300pg/ul) of my suspended plasmid solution in T.
- Waiting 30 min in ice
- Heat shock 30 sec 42°C
- Then 5min in ice
- Add 200uL SOC medium
- Recovery 2 hours then plate it on 3 different 100ug/mL Ampicillin LB
- Incubate 37°C till tomorrow
- I took 2 different colonies and do a 8 hours culture in two different tubes T1 an T2 in LB + Amp 100ug/mL, in order to do a glycerol stock.