Team:Kent/Experiments


iGEM Kent 2015

Experiments & Protocols

Contents

Competent Cells
Transformation Protocol
Miniprep

Competent Cells
Overview
Materials
Procedure
References
Transformation Protocol
Overview
Materials
Procedure
References
Miniprep
Overview

  • The Miniprep is for purification of molecular biology grade plasmid DNA
  • This provides a rapid method to purify plasmid DNA using silica membrane column

    • Materials
    Procedure

  • Add the provided RNase A solution to Buffer P1.
  • Mix the solution and store at 2–8°C
  • Add ethanol (96–100%) to Buffer PE before use
    1. Pellet 1–5 ml bacterial overnight culture by centrifugation at >8000 rpm (6800 x g) for 3 min at room temperature (15–25°C).
    1. Resuspend pelleted bacterial cells in 250 μl Buffer P1 and transfer it to a microcentrifuge tube.
    1. Add 250 μl Buffer P2 and mix thoroughly by inverting the tube 4–6 times until the solution becomes clear.'' Do not allow the lysis reaction to proceed for more than 5 min. If using LyseBlue reagent, the solution will turn blue.''

    References