Jessica Karen Wong/Notebook/2007-6-29

To Do

 * Digest E0240 with Mfe1 and Nsi1
 * PCR Clean E0240
 * Ligate E and 3K3, transform?
 * Colony PCR green T, and redo blue C gel
 * Miniprep B0032?

Overnights

 * B0032 has good growth
 * Also saw colonies on green T (promoter tester on Tet)

Digest

 * NEB recommends not double digesting Mfe1 and Nsi1
 * Doing a sequential digest Mfe1 then Nsi1
 * used 3ul of the scarred E0240 in the Mfe1 digest and all of the digested product in the 2nd (Nsi) digest
 * Mfe1 requires buffer 4 and Nsi1 requires buffer 3
 * Did a PCR cleanup in between the two digests

Colony PCR

 * Did a colony PCR on the 2 green (promoter tester) T colonies
 * Redid the colony PCR on the 7 blue (RBS tester) C colonies
 * Used VF and VR 40 uM primers
 * Made new 2.5uM dNTP's