Jessica Karen Wong/Notebook/2007-7-24

I2056 Scar/Promoter Gel

I2056

• Ran a gel of the overnight PCR
• Looks good
• Digesting with Mfe and Nsi in buffer 2 overnight

T9002

T9002 E0240 Gel

• Got new T9002_E0240_R primer in - diluted to 40 uM
• Set up a 100ul supermix PCR rxn of E0240 for T9002 to put on tails
  • Ran at 52
• Ran a gel which looks great
• PCR cleaned
• Digesting w/ Spe/Nsi in buffer 2 overnight

Other

• Made a new overnight from the registry plate of 1AK3
• Got sequencing back for I2055-3K3 - really wierd
  • 1st half of part (till middle of GFP) was put in backwards, rest of part was forwards
• Found that I2055 has an Mfe1 site in the middle of GFP and that's where the problem happened
• Changed synthesis proposal so doesn't have the Mfe1 site in GFP - silent mutation