User:Matthewmeisel/Notebook/2006-6-12

From OpenWetWare

< User:Matthewmeisel
Revision as of 16:37, 10 July 2006 by Matthewmeisel (Talk | contribs)
(diff) ←Older revision | Current revision (diff) | Newer revision→ (diff)
Jump to: navigation, search

DNA nanostructure reaction PCR

Transformation of standard components

  • goal: insert three BioBrick plasmids (already containing BioBricks) into E. coli in order to amplify them
    • a positive control (E7104) with the T7 promoter upstream of GFP
    • the lac operon (R0010)
    • GFP (E0241)
  • transformation protocol:
  1. 1 μL each of E7104, R0010, E0241, and water into separate microcentrifuge tubes that each contain 30 μL of competent cells
  2. Incubated the tubes on ice for 20 min.
  3. Heat shocked the tubes at 42°C for 30 s.
  4. Added 200 μL SOC to each tube.
  5. Incubated, with shaking, at 37°C for 1 h.
  6. Plated the mixture on carbomycin plates and incubated at 37°C for 24 h.
Personal tools