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

Ligation

Protocol

Ligation Name	Vector Name	Volume Vector (µL)	Insert	Volume Insert (µL)
L1	D187		D186
Control L1	D187		-	-

Cloning of EnvZ* in pSB1A2

Transformation results

	control			insert / vector mass ratio
	transformation with pUC19	transformation without plasmid	ligation without insert	1,8 / 1	2,4 / 1	3,1 / 1
number of colonies	many	0	0	8	5	2
number of clones picked up for screening				4	2	2

Team:Paris/August 27

From 2008.igem.org

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