2.2.2.3.3.1. PCR reaction mix

PCRs use a 20 µl final volume and should be set up as follows for the quantification of either N. apis or N. ceranae using primers from Forsgen and Fries (2010) in Table 1. Set up one mastermix as below for quantification of N. apis for each sample and set up another mastermix as below for quantification of N. ceranae for each sample, using either NaFor or NcFor respectively.

SsoFastTM EvaGreen® Supermix (BioRad)   1 x
NaFor or NcFor primer                                0.4 µM
UnivRev primer                                           0.4 µM
H20                                                           6.4 µl
Sample DNA extract                                   2 µl (ca. 2 ng DNA)