Haynes Lab:Notebook/SynBERC Scholars 2013/2014/02/13

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Procedure Digital PCR Part 1 TNF A forward and reverse primers were diluted from 100uM to 10uM. Initial stock DNA (Concentration = 2.3mg/mL) was diluted several times to obtain 4 different templates, listed below: U20S 1:100 = 6.24ng/mL D1 = 0.624ng/mL D2 = 0.0624ng/mL = 6.24*10^(-2) ng/mL D3 = 6.24pg/mL For each PCR reaction, the following volumes will be added: 1uL of forward TNF A primer 1uL of reverse TNF A primer 1uL of diluted DNA (either U20S 1:100, D1, D2, or D3 for each sample) 50uL of PCR master mix 47 uL of dH20 8 samples of each template will be put through the PCR sequence (NOTE: The program to be run is named "First Trial 121713") 2 samples of plain water (instead of template) will also be tested as a negative control.