Team:University College London/Protocols/Week14/1

From 2012.igem.org

Nuclease experiment for our nuclease BL21 + T7 promoter

For BL21 cell line which has inserted nuclease - Prepare 11-16 plates (10ml LBAgar+10ul CMP ).

- Streak cells onto all plates at the same time

- Incubate at 37°C

- Apply hydrochloric acid (HCL) to the first plate before putting them in the incubator (set time as zero)

- Take a second reading after four hours, followed by six readings every 3 hours, and a final three readings every two hours.

- When the reading is taken, observe the following:

a) Diameter of the colony (once the diameter of the colony is measured, pick the colony and put it to grow in LB for nine hours)

b) Diameter of the halo that is achieved once HCL is applied

c) OD from a)

d) Estimate of the depth of the colony on the agar plate