20.109(S08):Site-directed mutagenesis (Day2)

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20.109(S08): Laboratory Fundamentals of Biological Engineering

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Introduction

Protocols

Part 1: primer preparation

  1. Calculate the amount of water needed for each primer to give a concentration of 1 ng/mL.
  2. Touch-spin your primers, resuspend each in the appropriate volume of water, and touch-spin again.
  3. Calculate the dilution of primer that you need to prepare in order to have 125 ng present in 2.5 μl. Prepare 100-200 μL of this dilution and keep it on ice.

Part 2: site-directed mutagenesis

We will be using the QuickChange® kit from Stratagene to perform our site-directed mutageneses. Each group will set up three reactions and associated controls. You should work quickly but carefully, and keep your tubes in a chilled container at all times.

For next time

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