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From 2010.igem.org

Week #6

11th October - 15 October 2010

Monday

We kept inoculated cells on LB liquid medium at 37ºC overnight, each containing 30ml.

Prepared miniprep for I20260 and E0240, we are going to work on the characterize the promoter

and the cold expresion casstte by RPU's.

Tuesday

Yesterday's overnight inoculum we did miniprep and double digestion with EcoRI and PstI

To confirm the ligation of PCR 4.

DNA	10μl
Tango	1.8μl
EcoRI	1μl
PstI	1μl
H2O	1.20μl
Total	15μl

Wednesday

We ran a agarosa's gel to extract from the band E0240 and I20260.

DNA	50μl
Tango	9μl
EcoRI	5μl
PstI	5μl
H2O	6μl
Total	75μl

We cut the agarosa's band and purified with the kit.

At the same time we ran the PCR's with EcoRI and PstI

DNA	20μl
Tango	3.6μl
EcoRI	2μl
PstI	2μl
H2O	2.4μl
Total	30μl

Thursday

We Quantified with nano espectrophotometer the concentrations of PCR's products and the

Band's purified

	ng/μl
I20260	1.3 ng/μl
E0240	3.8 ng/μl
PCR 1 E and P	277.0 ng/μl
PCR 2 E and P	257 ng/μl
PCR 3 E and P	284.4 ng/μl
PCR 4 E and P	284.7 ng/μl