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. Make Ampicillin Plates