Folsomia candida, field population – replicate 1-Sanger
Springtails were sampled from a field location in Zaandam, the Netherlands, and transferred to standardized culturing conditions in the lab. After four months, DNA was isolated from four replicate samples, each consisting of ten animals, and used for 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.