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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 09:06, 15 September 2016
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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 [[ | # 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 [[McClean: designing primers]] 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. | # The desired end product will be this, where NNNN is the type specific overhang and can be found in the paper's SI. | ||
* Forward primer ---> | |||
* 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' | ||
* . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . <--- Reverse Primer | |||
# Step | # Step | ||
# Step 3 | # Step 3 | ||
