Methods
Summary
Molecular amplification with high fidelity polymerase and purification of DNA products provides the templates for sequencing of single species. Extra barcoding allows the metagenomic sequencing of many templates at once. A bioinformatics pipeline developed by the lead of the BeeMoS project at Hackuarium, for the OpenFoodRepo DNA analyses, WIMF (what's in my food) allows the sequence analyses of data from the MinION sequencing runs. The growth of mycelia on substrates for biomaterials (one of the earliest Hackuarium projects made lampshades of such biomaterials) will provide inserts for testing in beehives...
Challenges
Some challenges include sample status - pure or mixed, fresh or dry. We are already working on extraction methods to make dry samples more successful for amplification of the ribosomal DNA target used for species IDs. Another big challenge is making sure to have only one amplification product in the Sanger reactions. If two organisms are amplified, the sequence will provide very little of interest! Unless, of course, the plan is to sequence many barcoded templates at once, as we plan with the MinION again!
Pre Analysis Plan
Getting people involved to 'do it together' means more people can input and check the data in different ways. From our previous work, we know not only the benefits but the challenges and aim to address them. We will update all this with more links and info!
Protocols
This project has not yet shared any protocols.