User:JeffreyLau/Notebook/2006-6-15

2006/06/15
Ligation
 * Since our gel from yesterday showed failure, we ligated DNA extract from David and Peng.
 * 1) Mixed 6 µL insert (E0241), 2 µL vector (R0010), 2 µL DNA dilution buffer in reaction vial
 * 2) Added 10 µL ligation buffer
 * 3) Added 1 µL ligase, mixed
 * 4) Let incubate 5min at room temperature

Transformation
 * 1) Mixed 20 µL of ligation product with 30 µL competent cells
 * 2) Negative control: 2 µL of dH20 with 30 µL competent cells
 * 3) Positive control: 5 µL of positive control with 30 µL competent cells
 * 4) Incubate on ice for 20min
 * 5) Heat shock @42C for 30s
 * 6) Rest on ice for 2min
 * 7) Incubate overnight @37C