Registry/Measurement kit/Notebook/2007-7-26

Plates

• No growth on I2056.
• F2620-3K3 sample 1 and F2620-1AK3 have a few small colonies. F2620-3K3 sample 2 has 1 colony

To Do

• PCR clean digests from yesterday
• Ligate them
• Mini-Prep and digest 3K3 (E/X)

ccdB

• PCR products are digested and cleaned up.

I2055

• Ran gel of PCR, made o/n of only colony that looked right size.

T9002

• Running pcr of F2620-3k3 and 1AK3

BB PCRs

• Primers came today, so started E0240 (E-S) and I2057 (E-X) (Tm=52 deg)

Ligations

• I2055-1AK3 and 3K3
• I2056-1AK3 and 3K3
• I2057-1AK3
• E0240-1AK3

F2620-1AK3

I2055 Synthesis

• Decided not to make the silent point mutation; will use an AatII site on 3K3 (~100bp upstream of the cloning site) instead of MfeI