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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 09:23, 15 September 2016
, 15 September 2016→Protocol
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==Protocol== | ==Protocol== | ||
# Design primers as normal such that they can be used to PCR amplify your gene of interest. NCBI's Primer-Blast tool is handy for this. See [[McClean: designing primers]] for more help. | # Design primers as normal, such that they can be used to PCR amplify your gene of interest. NCBI's Primer-Blast tool is handy for this. See [[McClean: designing primers]] for more help. | ||
# Prefix the 5' end of your forward primer with GCAT'''CGTCTC'''ATC'''GGTCTC'''ANNNN | # Prefix the 5' end of your forward primer with GCAT'''CGTCTC'''ATC'''GGTCTC'''ANNNN | ||
# Prefix the 5' end of your reverse primer with ATGC'''CGTCTC'''A'''GGTCTC'''ANNNN | # Prefix the 5' end of your reverse primer with ATGC'''CGTCTC'''A'''GGTCTC'''ANNNN | ||
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* 3'- CGTAGCAGAGTAGCCAGAGTNNNN- gene of interest - NNNNA'''CTCTGG'''A'''CTCTGC'''CGTA -5' | * 3'- CGTAGCAGAGTAGCCAGAGTNNNN- gene of interest - NNNNA'''CTCTGG'''A'''CTCTGC'''CGTA -5' | ||
* . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . <--- Reverse Primer | * . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . <--- Reverse Primer | ||
==Notes== | ==Notes== | ||
