Jessica Karen Wong/Notebook/2007-7-7


 * Took out overnight transformation plates
 * both the sequential and double digests of T9002 and 3K3 had lots of colonies
 * E0240-3K3 had lots of colonies, E0240-1AK3 had two
 * I2055-1AK3 had lots of colonies, I-1AT3 and I-1AC3 had none


 * Made cell suspensions of 4 T-3K3 colonies, 4 I-1AK3 colonies, 4 E-3K3, and 2 E-1AK3
 * Did a 10 ul colony PCR on each cell suspensions
 * Used 2min elongation time because T9002 is 2261bp b/t VF and VR


 * Ran gel of I2055 and got a bright band just under 1kb
 * Not sure if it contains promoter or not
 * PCR cleaned I2055
 * Double digested I2055 with Mfe1 and Nsi1
 * Used buffer 2 and 4ul DNA


 * Overnight liquid cultures of I2056 still didn't grow (maybe lost Amp resistance?)
 * Made new overnights of I2056 containing only Tet


 * Did a 2-way ligation of T9002 and 1AK3
 * Transformed 3ul of the ligation and plated on Kan