Gene amplification from E. coli genome

LAB PROTOCOL

  1. Culture E. coli MG1655 strain overnight in LB medium

  2. Next morning, remove 100 microlitters of E. coli culture, place it in an eppendorf tube and boil for 10 minutes at 100 degrees Celcius. 

  3. Following, spin down boiled culture at 13,000 rpm for 30 second to remove cell debris.

  4. Remove 1 microlitter from supernatant and add this to the pre-prepared PCR reaction mix.

  5. Carry out PCR as recommended from the manufacturer of polymerase enzyme.


(Alternatively, if the E. coli MG1655 strain has been plated, cells from the colony can be added directly into the pre-prepared PCR mix. To break cells open, add a 10 minute boiling step at 100 degrees Celsius at the start of the PCR protocol.)

 
  • LinkedIn
  • Twitter

©2017