Team:Newcastle/4 August 2010

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Contents

Cloning the rocF BioBrick

Aim

The aim of today's experiment is to amplify 6 different fragments for the construction of rocF BioBrick with the help of 6 different Phusion PCR.

Materials and Protocol

Please refer to PCR for Phusion PCR protocol. The details for the 6 PCR reactions are mentioned below:

PCR

Tube Part to be amplified Plasmid consisting the part Forward primer Reverse Primer Melting Temperature (Tm in °C) Extension time* Size of the fragment
44.0 µl/ml 19.9 µl/ml 25.0 µl/ml 30.8 µl/ml 10.0 µl/ml 44.2 µl/ml 9.2 µl/ml 39.7 µl/ml

Gibson assembly

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