Team:TU Delft/27 juli 2010 content

Digestion

Some BioBricks are in production. Digestion reactions:

#	Sample	Enzyme 1	Enzyme 2	Enzyme 3	Buffer	Needed fragment
1	1.5 μg AlkS in pSB1C3	XbaI	SpeI		Buffer 2 + BSA (BioLabs)	’X - AlkS - S’
2	1.5 μg AlkS in pSB1C3	XbaI	SpeI	NCoI	Buffer 2 (BioLabs)	’X - AlkS - S’
3	1.0 μg B0015 in pSB1AK3	EcoRI	XbaI		Buffer 2 (BioLabs)	’E X’- B0015-pSB1C3
4	1.2 μg E0422 in pS-B1A2	EcoRI	XbaI		Buffer 2 (BioLabs)	‘E X’- E0422-pSB1C3

Results of the digestion on 1% agarose gel

1% Agarose of digestion check, gel runned at 100V for 1 hour, loaded 4 μL of marker and 10 μL sample + 2 μL loadingbuffer

Lane description:

#	Description	Expected Length (bp)	Status
1	Marker
2	Undigested B0015 in pSB1AK3	3318	✓
3	Digested B0015 in pSB1AK3 + EcoRI + XBaI	3303, 15	✓
4	Undigested E0422 in pSB1A2	2996	✓
5	Digested E0422 in pSB1A2 + EcoRI + XBaI	2981, 15	✓
6	Undigested AlkS in pSB1C3	4999	✓
7	Digested AlkS in pSB1C3 + EcoRI + SpeI + NCoI	2673, 1249, 1077	✓
8	Digested AlkS in pSB1C3 + EcoRI + SpeI	2673, 2326	✓

Conclusion: All the plasmids were digested as expected