Jessica Karen Wong/Notebook/2007-8-2

Sequencing Redos
Made overnights of 4 new colonies of I2056 (#4-7), I2057-ES, I2057-EX
 * I2057-EX-VR failed again
 * I2057-ES-VF failed again
 * E0240-ES-VF forward is perfect until 260, all is fine except for the 1bp mutation around 700
 * I2056-2-F still has the 1bp insertion at around 20, rest is fine
 * I2056-3K3-1F has a 1bp deletion at about 60, rest is fine
 * I2055-3K3-1-F perfect until 758, full coverage and scarred

I2055

 * PCR cleaned overnight digest and RBS digests (B0030, 31, 32)
 * Has complete go ahead
 * Ligated:
 * I2055-NOT-EX closed
 * I2055-Not-SX closed
 * I2055-EX-B0032
 * I2055-SX-B0032
 * I2055-SX-B0030
 * I2055-SX-B0031

I2056 - overnight plates had no growth, seq was bad, will be sending in colonies 4-7 to seq

I2057

 * Minipreped I2057-ES and I2057-EX and digested E/S and E/X respectively
 * Didn't get enough DNA so making another overnight to use
 * Also overnighting to seq
 * Digested I2057-ES that had been sealed with Not1, cut E/S
 * Ligated PCR product cut either ES and EX w/ promoters
 * I2057-ES-R0040
 * I2057-EX-R0040
 * I2057-EX-J100
 * I2057-EX-J116
 * PCR cleaned I2057-ES E/S digest from today

Made new Kan plates

F2620

 * Ran gel of overnight PCR - looked good
 * PCR cleaned
 * F2620-3K3 plate from overnight had no growth
 * Digested PCR product Mfe/Xba in buffer 4
 * PCR cleaned digest