User:Anthony Salvagno/Notebook/Research/E. coli Transformation Protocol

Today we are inserting the plasmids into E. coli cells. Here is the protocol:

Prep
For general protocol modify steps 1, 2, and 3 per user discretion.
 * 1) Need 10ng/uL plasmid DNA
 * 2) *from a 527ng/uL stock I wanted 200uL stock of above concentration. That means I needed to add 3.8uL of plasmid DNA to 196uL TE buffer.
 * 3) Create 4 tubes
 * 4) No DNA
 * 5) 2.5uL DNA (from 10ng/uL stock)
 * 6) 5uL DNA
 * 7) 10uL DNA
 * 8) combine with 50uL competant E. coli cells.
 * 9) Incubate on ice for 30 min
 * 10) Heat shock for 1 min at 42C
 * 11) Add 1mL LB
 * 12) incubate for 60 min at 37C in shaker
 * 13) Spin cells at 4krpm for 1 min
 * 14) discard ~850uL supernatant
 * 15) Mix cells in remaining 150uL
 * 16) plate on LB and amp using glass beads
 * 17) Incubate plates at 37C overnight