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From 2010.igem.org

Alkane degradation

Today another DLT - we will stick a strong promoter (J13002) in front of ladA

#	Sample	Enzyme 1	Enzyme 2	Enzyme 3	Buffer	BSA	Needed fragment
1	LadA	XbaI	PstI	AseI	3 (Biolabs)	✓	‘X – ladA – P’
2	J13002	SpeI	PstI	✗	3 (Biolabs)	✓	‘P – pSB1A2 – J13002 – S’

Directly following the digestion, the products were ligated for 1 hour:

#	BioBrick	Fragment 1	Fragment 2
1	pSB1A2 - J13002 - ladA	‘X – ladA – P’	‘P – pSB1A2 – J13002 – S’