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Team:Cambridge-JIC/Guide/Constructs/MoCloAssembly
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For protocols and outline of MoClo, see [http://pubs.acs.org/doi/pdf/10.1021/sb4001504 "Engler et al.:A modular cloning toolbox for plant cloning] or [[RFC 105]]. | For protocols and outline of MoClo, see [http://pubs.acs.org/doi/pdf/10.1021/sb4001504 "Engler et al.:A modular cloning toolbox for plant cloning] or [[RFC 105]]. | ||
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3. Clone them into the appropriate acceptor. | 3. Clone them into the appropriate acceptor. | ||
See RFC105 for further details. | See RFC105 for further details. | ||
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Revision as of 15:26, 17 October 2014
MoClo Assembly
For protocols and outline of MoClo, see [http://pubs.acs.org/doi/pdf/10.1021/sb4001504 "Engler et al.:A modular cloning toolbox for plant cloning] or RFC 105.
If using MoClo, it is recommended that you use the [http://parts.igem.org/Part:BBa_K1484316 selection casette] as the first of your level 1 parts.
After cloning into a level 1 construct, and amplifying the plasmid in E-Coli, it is suitable for direct electroporation into agrobacteria
Parts from other sources
If you're using Level 0 parts not from MoClo plasmids, you need to:
1. Make sure they don't have Bsa1, Bpi1 and BsmBI restriction sites
2. PCR Amplify them using primers:
Forward: AAAAGGTCTCANNNN_your_binding_forward_primer
Reverse: AAAAGGTCTCA[MMMM]'_your_binding_reverse_primer,
where NNNN denotes the according PlantSyntax/RFC105 code for the 5' end of the part, and [MMMM]' the reverse complement of the code for the 3' end.
3. Clone them into the appropriate acceptor.
See RFC105 for further details.
