PCR Back on Track

After some issues with primers we are back on track. Each pillbug sample will have 5 PCR runs. That's a lot of PCR. From these, three potentially can be sent for sequencing. The COI will be sent for mitochindrial comparison and check on species ID. The Wsp primers will help us identify the presence of Wolbachia and the supergroup. If we have evidnece of the f-element nuclear insert, that will be sent also. We have not seen the f-element much in the US populations so we will extract DNA fom gel bands and sequence those to confirm the correct match. So, a lot of molecular work on each pillbug. Thus far we have extracted DNA from 152 pillbugs from different geographic locations. We normally use head tissue but we have found that pervorming an additional purification step to the extraction is goiving us much better PCR results. Stay tuned for another update soon!