Team:Newcastle/4 August 2010

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Line 18: Line 18:
!'''Reverse Primer'''
!'''Reverse Primer'''
!'''Melting Temperature (Tm in °C) '''
!'''Melting Temperature (Tm in °C) '''
-
!'''Size of the fragment'''
+
!'''Size of the fragment (in bp)'''
-
!'''Extension time*'''
+
!'''Extension time* (in seconds)'''
|-
|-
-
|44.0 µl/ml
+
|1
-
|19.9 µl/ml
+
|Plasmid Vector
-
|25.0 µl/ml
+
|pSB1C3
-
|30.8 µl/ml
+
|P1V1 forward
-
|10.0 µl/ml
+
|P2V1 reverse
-
|44.2 µl/ml
+
|58
-
|9.2 µl/ml
+
|2072 approx.
-
|39.7 µl/ml
+
|60
|}
|}

Revision as of 09:17, 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) Size of the fragment (in bp) Extension time* (in seconds)
1 Plasmid Vector pSB1C3 P1V1 forward P2V1 reverse 58 2072 approx. 60

Gibson assembly

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