Jessica Karen Wong/Notebook/2007-7-6

To Do

 * Redo E0240 PCR w/ vent and phusion
 * Ligate, transform T9002 and 3k3
 * Remake overnight cultures of I2056

E0240

 * Redid 2 100ul prep pcr's with Vent and Phusion
 * Just realized that we completely skipped the ligation and transformation with backbone
 * Discarded PCR product
 * Did two way ligations E0240-3K3 and E0240-1AK3 (used 4.5ul of each E0240 and backbone)
 * Transformed 3 ul of each ligation and plated

T9002

 * Ligated T9002-D and T9002-S with 3K3 (Double and Sequentially digested)
 * Transformed using 3ul of ligation
 * Plated on Kan plates

I2055

 * Got sequencing back and all samples do not contain R0040
 * Religated R0040-I13401-1AK3, R-I-1AT3, R-I-1AC3 (3ul each part)
 * Transformed 3 ul of each ligation and plated
 * Primer to PCR in R0040 came in
 * Doing a 100ul prep PCR of the #4 colony of I2055 on 1AK3
 * Using Vent at 53C (1:30 extension time)

From now on should use Vent for all Preparatory PCR reactions

I2056

 * Overnight liquid cultures did not grow
 * Made new overnights from actual colony instead of cell suspension