Team:Nevada/Protocols

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(Difference between revisions)
(Gibson Assembly)
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==Gibson Assembly==
==Gibson Assembly==
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:::#1.Please note that this protocol has been optimized for the assembly and cloning of one or two fragments into any vector. For the cloning of larger assemblies, we recommend using the standard protocol provided with the Gibson Assembly Master Mix
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:::#1.Please note that this protocol has been optimized for the assembly and cloning of one or two fragments into any vector. For the cloning of larger assemblies, we recommend using the standard protocol provided with the Gibson :::#Assembly Master Mix
Gibson Assembly Reaction Setup
Gibson Assembly Reaction Setup
Amount per Reaction
Amount per Reaction
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:::#3.Incubate samples in a thermocycler at 50°C for 15 minutes. After incubation, store the samples on ice or at -20°C for subsequent transformation.
:::#3.Incubate samples in a thermocycler at 50°C for 15 minutes. After incubation, store the samples on ice or at -20°C for subsequent transformation.
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Note: Extended incubation up to 60 minutes may help to improve assembly efficiency in some cases (for further details see FAQ section).
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:::#Note: Extended incubation up to 60 minutes may help to improve assembly efficiency in some cases (for further details see FAQ section).
:::#4.Transform NEB 5-alpha Competent E. coli cells (provided with the kit) with 2 μl of the assembly reaction, following the transformation protocol.
:::#4.Transform NEB 5-alpha Competent E. coli cells (provided with the kit) with 2 μl of the assembly reaction, following the transformation protocol.

Revision as of 04:20, 28 September 2013