Screening of the cloning of E0240 and FlhDC+promotor

Spreading the clones in order to obtain single colonies

Strain	Resistance	Ligation	DNA cloned	vector	expected size of the fragment amplified by VF & VR	mesured size
S159.1	kanamycine	L139.1	E0240 (GFP tripart)	pSB3K3	1192 bp	1,5 kb 1,1 kb 0,6 kb
S161.1	ampicilline	L142.7	FlhDC+promotor	pSB1A2	1403 bp	1,4 0,4 kb 0,3 kb

The plates obtained from the speading of yesterday can't be used because there are not single colonies.
We have to try again, but with a stronger dilution of the bacteria or with a smaller volume of spreading.

Digestion

Aim : Construction of "pFlgA-RBS-YFP-dbl ter"

Measurement of concentration of minipreps

standard protocol

Miniprep	Biobrick	C° (µg/mL)	ratio 260/280
MP164.1	E0422	95	1.69
MP164.2	E422	90	1.76
MP165.1	E0430	131	1.74
MP165.2	E0430	nd	nd
MP166.1	E0432	112	1.65
MP166.2	E0432	79	1.6
MP167.1	E0420	199	1.72
MP167.2	E0420	194	1.73
MP168.1	I732078	111	1.65
MP168.2	I732078	104	1.67
MP122.1	E0840	56	1.58
MP122.2	E0840	98	1.63

Digestion

Protocol Digestion

Name	Template DNA	Description	Vol MP (µl)	Vol H2O (µl)	Enzymes
D166	MP165.1	- FV	7.63	17	EcoRI and XbaI
D167	MP166.1	- BV	8.9	15.8	EcoRI and XbaI
D131	MP122.2	- BV	10.2	14.5	XbaI and PstI

Team:Paris/August 20

From 2008.igem.org

Contents

Screening of the cloning of E0240 and FlhDC+promotor

Spreading the clones in order to obtain single colonies

Digestion

Measurement of concentration of minipreps

Digestion