Sensor: Week 6 June 20-25

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Revision as of 18:01, 2 July 2011

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 O_R 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+O_R 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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 |-
 |Transformation/Plating
-|colspan="2"|The above ligations and K3+O<sub>R</sub> were transformed into Escherichia coli cells. All cells were plated on plates with appropriate resistance.
+|colspan="2"|The above ligations and K3+O<sub>R</sub> were transformed<br />into Escherichia coli cells. All cells were plated<br />on plates with appropriate resistance.
 |}