2.4.2. DNA amplification
DNA amplification requires a PCR machine (such as an Eppendorf Mastercycler®) and a set of specific forward and reverse primers. The machine is initially programmed to carry out a number of cycles to amplify the DNA (see the section ‘DNA methods’ of the BEEBOOK paper on molecular methods (Evans et al., 2013)).
A commonly used method for amplifying Varroa spp. DNA consists of:
1. PCR thermo-cycles of 5 min pre-denaturation at 94 oC.
2. 35 cycles of denaturation at 94 oC for 1 min.
3. 1 min annealing at 52 oC.
4. 2 min extension at 72 oC.
5. Final extension at 72 oC for 5 min.