In the last two qPCR assays, no fluorescence was detected. Turns out we were using Promega master mix which is specifically for probe-based assays…as such, it doesn’t have any fluorescent dye in it, explaining the lack of fluorescence detection in any of the assays. I switched over to using a different master mix that should solve the issue.
I reran the DNMT1 primer assays and will run an actin qPCR to determine if it might be a possible normalizing gene. For protocol used, please refer to previous lab book entries.
Link to Excel spreadsheet of Cq values for both actin and DNMT1: https://docs.google.com/spreadsheets/d/1AhdJLEcXt6vnNBcGVLnc926KlZ5J-Nt8JMpiCdrUvaU/edit?usp=sharing
Link to DNMT1 qPCR data: http://owl.fish.washington.edu/scaphapoda/qPCR_data/cfx_connect_data/admin_2018-12-12%2015-24-28_BR006896.pcrd
Link to actin qPCR data: http://owl.fish.washington.edu/scaphapoda/qPCR_data/cfx_connect_data/admin_2018-12-14%2016-16-16_BR006896.pcrd
Plate map:
All replicates are in adjacent wells. The order of the samples in the well plate by rows is: D01, D02, D03, D04, D05, D06, D07, D08, D09, D10, D11, D12, D13, D14, D15, D16, D17, D18, D19, D20, T01, T02, T03, T04, T05, T06, T07, T08, T09, T10, T11, T12, T13, T14, T15, T16, T17, T18, T19, T20
Example:
Well—–Sample
A01——D01
A02——D01
A03——D02
A04——D02
Roberts Lab 