Team:Evry/Interlab Study/09-10-2014
From 2014.igem.org
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Construction n°1: PSB1C3 with I20260
Construction n°2: PSB1C3 with J23101-E1010
Construction n°3: PSB1C3 with K823012-E1010
Other constructions of the Anderson library of constitutive promoters
Miniprep disgestion of PSB1C3+E0240 plasmid.
- 1 µl EcoRI
- 1 µl XbaI
- 2 µl buffer 2.1
- 0.2 µl BSA
- 5 µl template (miniprep)
- 10.8 µl miliQ watter
Miniprep disgestion of BBa_J61002+J23101, BBa_J61002+J23118, BBa_J61002+J23105, BBa_J61002+J23106 and BBa_J61002+J23107 plasmids.
- 1 µl EcoRI
- 1 µl SpeI
- 2 µl buffer 2.1
- 0.2 µl BSA
- 5 µl template (miniprep)
- 10.8 µl miliQ watter
Ligation to obtain constructions PSB1C3+E0240+J23101, PSB1C3+E0240+J23118, PSB1C3+E0240+J23105, PSB1C3+E0240+J23106 and PSB1C3+E0240+J23107.
- 1 µl T4 DNA ligase
- 6 µl Insert (digested BBa_J61002+J23101, BBa_J61002+J23118, BBa_J61002+J23105, BBa_J61002+J23106 and BBa_J61002+J23107)
- 2 µl vector (digested PSB1C3+E0240)
- 2 µl buffer
- 9 µl miliQ watter
Transformation in E. coli DH5 alpha with the protocol of the 15th of August. Bacteria were plated on Chloramphenicol agar plates and incubated at 37°C overnight.
Sep 10