Primer Design for qPCR: Difference between revisions
m added a first step of checking the primer availability in the lab using the primer data sheet in google drive before ordering the primer and edited the last step as to where to add the new primers (on the primer data sheet) |
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==Making Your Own Primers From Entrez== | ==Making Your Own Primers From Entrez== | ||
# Check if the primer is available in the lab using the Primer Data sheet at https://docs.google.com/spreadsheets/d/1kQ1G52XuoqpAWHIWT5H2UDz4aDhP285oUyYcj7XTB24/edit?ts=5925e7a2#gid=0. If the primers are not available, proceed to the next steps | # Check if the primer is available in the lab using the Primer Data sheet at https://docs.google.com/spreadsheets/d/1kQ1G52XuoqpAWHIWT5H2UDz4aDhP285oUyYcj7XTB24/edit?ts=5925e7a2#gid=0. If the primers are not available, proceed to the next steps | ||
#Find your RNA sequence on entrez by starting with the gene at http://www.ncbi.nlm.nih.gov/gene (make sure its the correct species). Then pick the mRNA you want to probe, based on the isoform structure on that page. Click on that nucleotide | #Find your RNA sequence on entrez by starting with the gene at http://www.ncbi.nlm.nih.gov/gene (make sure its the correct species). Then pick the mRNA you want to probe, based on the isoform structure on that page. Click on that nucleotide. (it will be an NM number) | ||
# Under analyse this sequence click '''Pick Primers''' | # Under analyse this sequence click '''Pick Primers''' | ||
# Under PCR Product Size pick 70-150 as the range | # Under PCR Product Size pick 70-150 as the range | ||