Sample information

Wolbachia positive fruit fly from The Wolbachia Project

Methods

Duplex PCR was performed

Results

The purpose of this experiment is to isolate DNA from organisms and determine if there is Wolbachia DNA present by using PCR. Arthropod samples were lysed and the DNA was extracted from the samples. PCR was used to amplify the CO1 gene found in arthropods and the W16S gene found in Wolbachia. The CO1 gene has a length of 708 base pairs and the W16S gene has a length of 438 base pairs. There were 5 samples that were run through PCR: a sample, +Wolbachia, -Wolbachia, a +DNA control, and a -DNA control (water).  The +Wolbachia and the +DNA control were expected to have two bands, one at 708 BP (CO1) and one at 438 BP (W16S). As seen in Figure 2, well 4 (+Wolbachia) and well 6 (+DNA control) had two bands as expected. The -Wolbachia sample was expected to have one band at 708 bp. As seen in Figure 2, well 5 (-Wolbachia) had one band as expected. The -DNA control (water) was expected to have no bands as it should not contain DNA. As seen in Figure 2, well 7 (-DNA control) did not have a band. Lastly, the sample should have either one or two bands, depending on if the arthropod was infected. As seen in Figure 2, well 3 (sample) had one band at 708 bp and one at 438 bp, meaning that the sample was infected with Wolbachia. Due to the controls having the expected results, the experimental results are valid. It can be said with high confidence that the sample was infected with Wolbachia.