From 2010.igem.org
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)
| Size of the fragment
| Extension time*
|
| 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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