Erickson:REal-time PCR

Isolation of Total RNA from Fleas
Master Mix: •	5μL 2x SYBR green •	1μL Forward primer (5μM) •	1μL Reverse primer(5μM) •	3μL PCR grade water
 * Note: This is for a single reaction, multiply by number of reactions needed

Procedure  1. Add 1μL of DNA or cDNA teplate to 9μL Master Mix in a 96-well Multiwell plate. 2. Seal the multiwell plate with Light Cycler 480 Multiwell sealing foil 3. Centrifuge the Multiwell plate at 15xg for 2 minutes 4. Transfer the Multiwell plate to the holder of the Light Cycler and close(push button to open and close) 5. Start the PCR program