Folsomia candida, laboratory population – replicate 3-Sanger
Springtails were sampled from a population kept in standardized culturing conditions for several years. Four replicate samples were collected, each consisting of ten animals, and used for DNA isolation and targeted PCR. A nested approach was used to suppress amplification of Wolbachia DNA: first, a PCR was performed with primers 8f and 1525R, and the resulting PCR products were amplified with primers 357F and 1392R. The DNA fragments were then purified, inserted into plasmid pGEM-T and transformed into Escherichia coli XL-1 Blue. Positive transformants were screened by PCR to verify the insertion of the fragment and then subjected to RFLP analysis. Six to eight clones showing different RFLP patterns were chosen for sequencing.