Team:TP-CC San Diego/Protocols


Protocols

TRANSFORMATION PROTOCOL

Procedure:

  1. Thaw comp cells on ice for 10 mins.
  2. Add 10uL of DNA to comp cells.
  3. Let sit on ice for 10 mins
  4. Heat shock at 42˚C for 1 min
  5. Let sit on ice for 2 min
  6. Add 300uL of SOC media
  7. Put in 37˚C shaker at 200 rpm for 1 hr
  8. Place agar plates in 37˚C incubator to warm up; remove when needed.
  9. Pipette 300uL of comp cells onto agar plate
  10. Use sterilized beads to spread comp cells evenly
  11. Place in 37˚C incubator overnight

INOCULATION PROTOCOL

Procedure:

  1. Add 5mL of LB to Polypropylene round bottom tube
  2. Add 5uL of Carb antibiotic to LB
  3. Label and pick colony from plate
  4. Loosely secure cap to allow air flow
  5. Place in 37˚C shaker at 200rpm overnight

GUIDE RNA DESIGN

Procedure:

  1. Add 5mL of LB to Polypropylene round bottom tube

DIGESTION PROTOCOL

Procedure:

  1. Add 5mL of LB to Polypropylene round bottom tube

ANNEALING PROTOCOL

Procedure:

  1. Add 5mL of LB to Polypropylene round bottom tube

LIGATION PROTOCOL

Procedure:

  1. Add 5mL of LB to Polypropylene round bottom tube

SPIN PROTOCOL

Procedure:

  1. Add 5mL of LB to Polypropylene round bottom tube