Team:Newcastle/4 August 2010

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{|border=1
{|border=1
|-
|-
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!'''pSB1C3
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!'''Tube'''
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(No. 1)'''
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!'''Part to be amplified'''
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!'''pSB1C3
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!'''Plasmid consisting the part'''
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(No. 2)'''
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!'''Forward primer'''
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!'''pSB1C3
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!'''Reverse Primer'''
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(No. 3)'''
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!'''Melting Temperature (Tm in °C) '''
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!'''pSB1C3
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!'''Extension time*'''
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(No. 4)'''
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!'''Size of the fragment'''
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!'''lacI
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(No. 1)'''
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!'''lacI
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(No. 2)'''
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!'''Double terminator !'''Double terminator
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(No. 1)'''
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!'''Double terminator
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(No. 2)'''
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|44.0 µl/ml
|44.0 µl/ml

Revision as of 09:13, 5 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

Newcastle University logo.png    Newcastle cbcb logo.pngNewcastle Biomedicine logo.gif    Team Newcastle CEG logo.gif
Newcastle iww logo.jpg  UNIPV Pavia Logo.gif  Newcastle BBSRC.gif    Newcastle Genevision logo.png Newcastle WelcomeTrust.jpg
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