Temperature Cycling:

• 92 - 94^oC for 30 - 60 sec (denature)

• 37 - 72^oC for 30 - 60 sec (anneal)

• 72^oC for 30 - 60 sec (elongate) (60 sec per kb target sequence length)

• 25 - 35 cycles only (otherwise enzyme decay causes artifacts)

• 72^oC for 5 min at end to allow complete elongation of all product DNA

NOTE:

"Quickie" PCR is quite feasible: eg, [94^oC 30 sec / 45^oC 30 sec / 72^oC 30 sec] x 30, for short products (200 - 500 bp).

YOU CAN USE GLYCEROL IN THERMAL CYCLER REACTION TUBE HOLES TO ENSURE GOOD THERMAL CONTACTS

DON'T RUN TOO MANY CYCLES: if you don't see a band with 30 cycles you probably won't after 40; rather take an aliquot from the reaction mix and re-PCR with fresh reagents.  See here.