Jessica Karen Wong/Notebook/2007-8-1

I2057

• Transformation plates w/ DB3.1 had no colonies
• Doing a positive control on DB3.1 w/ puc18
• Was going to digest the preps of I2057-EX and I2057-ES resealed w/ Not1, but didn't have enough DNA
• Making a 10ml overnight culture of each
• Minipreped overnight cultures of promoters J23116 and J23100
• Digested R0040, J23116, and J23100 - Eco/Spe

I2056

• PCR cleaned overnight digests of I2056-EX w/ E/X, I2056-EX w/ Not, I2056-SX S/X, I2056-SX w/ Not1
• Ligated I2056-EX-N and I2056-SX-N each to themselves (closed up)
• Transformed in Top10 and plated
• Digested RBS's B0030, B0031, and B0032 E/S and X/S each for later testing

I2055

BB I2055 PCR

• Ran a gel of the overnight BB PCR's loaded: ladder sp I2055-EX sp I2055-SX
• Looked the right size
• PCR Cleaned overnight PCRs
• Digesting I2055-SX with S/X, I2055-SX w/ Not1, I2055-EX with E/X, I2055-EX w/ Not1 overnight

F2620

• Transformation plates had no colonies
• Religated F2620-3K3
• Transformed with Top10 and plated
• We're running out of DNA, so made new PCR
• Ran 1 100ul supermix PCR to add tails to F2620 in 1A3 at 53 w/ 1:30 ext time