Template Preparation for RT-qPCR
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Template Preparation
1. Place 1mL of grown bacteria into a 1.5mL tube.
2. Heat the tube at 95C for 5min in a water bath. (If tubes pop open frequently try reducing the volume to .5mL, and/or reinforce lids with tape or clip.)
3. Immediately after heating, place the tubes in a -20C cooler/freezer for 10min or until frozen.
4. Take out and allow to thaw for dilutions.
Dilutions of Prepared Templates
After prepared templates have thawed perform serial dilutions of 1:10, 1:100, 1:1000 making sure to vortex before each dilution.
-When using 1mL volume of template, dilute .1mL of template into .9mL of dH2O.
-When using .5mL volume of template, dilute .05mL of template into .45mL of dH2O.
