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McClean: Designing "Yeast Toolkit" compatible primers: Difference between revisions
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McClean: Designing "Yeast Toolkit" compatible primers (view source)
Revision as of 09:01, 15 September 2016
, 15 September 2016no edit summary
(New page: ==Overview== This protocol is written to prepare the user to design PCR primers such that the end product will have BsaI and BsmBI cut sites, as well as the correct flanking overhangs. It ...) |
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==Protocol== | ==Protocol== | ||
# | # Design primers as normal such that theycan be used to PCR amplify your gene of interest. NCBI's Primer-Blast tool is handy for this. See [[Link title]] for more help. | ||
# The desired end product will be this, where NNNN is the type specific overhang and can be found in the paper's SI. | |||
* 5'- GCAT'''CGTCTC'''ATC'''GGTCTC'''ANNNN- gene of interest - NNNNTGAGACCTGAGACGGCAT -3' | * 5'- GCAT'''CGTCTC'''ATC'''GGTCTC'''ANNNN- gene of interest - NNNNTGAGACCTGAGACGGCAT -3' | ||
* 3'- CGTAGCAGAGTAGCCAGAGTNNNN- gene of interest - NNNNA'''CTCTGG'''A'''CTCTGC'''CGTA -5' | * 3'- CGTAGCAGAGTAGCCAGAGTNNNN- gene of interest - NNNNA'''CTCTGG'''A'''CTCTGC'''CGTA -5' | ||
