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