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From 2010.igem.org

Alkane Sensing, Solvent Tolerance and Salt Tolerance

by Pieter

The plates containing yesterday's ligations contained colonies, to check whether they really contain the desired BioBrick a colony PCR was done, and the used colonies were grown in liquid LB medium over night. The results from the PCR were analysed on a 1% agarose gel.

Alkane degradation

For the next step in BioBrick formation a digestion was done:

#	Sample	Enzyme 1	Enzyme 2	Enzyme 3	Buffer	BSA	Needed fragment
1	1 μg 007T	EcoRI	SpeI	BamH1	2 (Biolabs)	✓	‘E – J61100-AlkB2 – S’
2	1 μg 008A	EcoRI	XbaI		2 (Biolabs)	✓	‘E – J61100-rubA3 – X’
3	1 μg 010A	EcoRI	SpeI	AseI	2 (Biolabs)	✓	‘E – J61100-rubR – S’
4	2 μg B0015	EcoRI	XbaI		2 (Biolabs)	✓	‘E – B0015 – pSB1AK3 – X’
5	1 μg 017A	EcoRI	SpeI	AseI	2 (Biolabs)	✓	‘E – J61100-ladA – S’
6	1 μg 018A	XbaI	PstI	AseI	2 (Biolabs)	✓	‘X – J61101-ADH – P’
7	1 μg 019A	EcoRI	SpeI	AseI	2 (Biolabs)	✓	‘E – J61107-ALDH – S’
8	1 μg pSB1K3	EcoRI	PstI		2 (Biolabs)	✓	‘E – pSB1K3 – P’

Results of the digestion on 1% agarose gel

gel runned at 100V for 1 hour loaded 5 μL of marker and 5 μL sample + 1 μL loadingbuffer