Cyanobacterial Primers (cyano)

First Cycle (run once)

• 95oC for 5 minutes
• 80oC for 1 minute
• With about 3 seconds remaining, pause thermocycler and add Taq

Second Cycle (run 30 times)

• 94oC for 1 minute
• 60oC for 1 minute
• 72oC for 1 minute

Third Cycle (run once)

• 72oC for 9 minutes
• 4oC hold

Eubacterial Primers (eubac td)

First Cycle (run once)

• 95oC for 5 minutes
• 80oC for 1 minute
• With about 3 seconds remaining, pause thermocycler and add Taq
• 65oC for 1 minute
• 75oC for 3 minutes

Second Cycle, Touchdown PCR (run 20 times)

• 94oC for 1 minute
• 65oC for 1 minute
• 75oC for 3 minute
• NOTE: this is a touchdown cycle; the temperature for the second step must be lowered by 1oC every second cycle (ending on 55oC).

Third Cycle (run 9 times)

• 94oC for 1 minute
• 55oC for 1 minute
• 72oC for 3 minute

Fourth Cycle (run once)

• 72oC for 7 minutes
• 4oC hold