Team:Paris/September 1

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Contents

1 Cloning of EnvZ*
- 1.1 Transformation results of L165 (EnvZ*-pSB1A2) from August 29th
2 Construction of pFlhB - mRFP Tripart (LVA+)
- 2.1 Digestion
  - 2.1.1 Measurement of the concentration of D187 purified
- 2.2 Ligation

Cloning of EnvZ*

Transformation results of L165 (EnvZ*-pSB1A2) from August 29th

sample	number of colonies
positive control pUC19	many colonies
transformation negative control	0
ligation without insert	0
3:1 insert:vector ratio	5
4:1 insert / vector ratio	5

Construction of pFlhB - mRFP Tripart (LVA+)

Aim : Construction of "pFlhB-RBS-mRFP-dbl ter" (pFlhB-I732078)
We can only do the construction with mRFP Tripart (LVA+) because the stable strain with the Biobricks I732011 (mRFP Tripart LVA-) don't to growth.

Digestion

Measurement of the concentration of D187 purified

Protocol (it's same that for Miniprep)

Digestion Name	Concentration (µg/mL)	Ratio 260/280
D187	16	1.31

Ligation

Ligation Name	Vector Name	Volume Vector (µL)	Insert	Volume Insert (µL)
L168	D187		D186
Control L168	D166		-

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