Erickson:REal-time PCR
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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