Minnesota/9 July 2008

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1. Single Digests: Performed today. Incubated digested samples for 2hrs @ 37C. Refer to table below.

Gene	10x Buffer	BSA Protein	H20	DNA	RE
Promoter	5.0uL	0.5uL	40.4uL	2.1uL	2.0uL, SPE
LAMBDAcI	5.0uL	0.5uL	29.5uL	15.0uL	1.0uL, XBA 1
LAMBDAcI	5.0uL	0.5uL	29.5uL	15.0uL	1.0uL, SPE
Terminator	5.0uL	0.5uL	38.0uL	4.5uL	2.0uL, XBA 1

NOTE: RE = Restriction Enzyme

2. Ligation: Performed today. Incubate ligated samples @ 16C for 1hour. Heat inactivated enzyme @ 65C for 15 min. Refer to table below.

Genes	10x Buffer	H20	Base Vector	Insert	DNA Ligase
LAMBDAcI/Terminator	2.0uL	10.7uL	2.0uL (Term.)	4.3uL (LAMBDAcI)	1.0uL
Promoter/LAMBDAcI	2.0uL	10.7uL	2.0uL (Pro.)	4.3uL (LAMBDAcI)	1.0uL

3. Double Digests

4. Make/Pour Ampicillin Plates