Sensor: Week 6 June 20-25

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Revision as of 18:00, 2 July 2011

Contents

Tuesday June 21

Restriction Enzyme Test II

The sensor group digested the K3 vector with the following restriction digests:

EcoRI
EcoRI XbaI
SpeI PstI
EcoRI PstI
XbaI SpeI

These digests were run on an agarose gel. The bands on the gel showed that the restriction enzymes were working.

Wednesday June 22

Radiation Sensor Construct, Day 1

The sensor group continued to assemble their test construct by placing a constitutive promoter in front of two RBS + cI repressor. The sensor group also attempted to place the OR site in front of the test construct pieces (K648000+K648001) and in front of an RBS and reporter. The sensor group tried to clone Cro into K3 and put an RBS in front of the cI repressor.

Assembly Step Part Involved with Assembly Step
Insert PCR J23113
B0034+mCherry
K648000
Restriction Digest Inserts using XbaI and PstI: B0034+mCherry
K648000+K648001
Cro
C0051
Insert using EcoRI and SpeI: J23113
Vectors using SpeI and PstI: K3+OR
B0032
Vector using EcoRI and XbaI: B0031+C0051
Vector using XbaI and PstI: K3+RFP
Ligation OR + B0034+mCherry
OR + K648000+K648001
J23113 + B0031+C0051
K3 + Cro
B0032 + C0051
J23113 + B0030+C0051
Transformation/Plating The above ligations and K3+OR were transformed into Escherichia coli cells. All cells were plated on plates with appropriate resistance.

B0034+C0051 and K3 were grown overnight in culture in order to extract more plasmid.




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