Culture Protocols and Information for T brucei Culture protocols and information for T brucei. Detailed guide to the practice and history ofTrypanosoma brucei cultivation. This covers the culture of bloodstream and procyclic forms in liquid media and on agarose plates.
Isolation of Live Bacteria from Adult Insects Protocol Protocol was developed to isolate Wolbachia from adult Drosophila, but it can be adapted for other insects. In some insects leg removal prior to isolation facilitates hemolymph extrusion.
Lysing Colonies and Binding of DNA to Filters Protocol Alkali is used to liberate DNA from bacterial colonies on nitrocellulose or nylon filters. The DNA is then fixed to the filter by UV-cross-linking or baking under vacuum.
Microbial Detection in Ice Tea Using the Millipore Milliflex Rapid Microbiology Detection System Ice tea has a complex composition, which leads to reduced filterability, and a decrease in sample throughput. Its composition can generate background or false positive signals. It is also well known that ice tea contains molecules that can inhibit the bioluminescence reaction, which can generate false negative results. The aim of this study was to develop a protocol that was able to neutralize these affects and enable faster detection of contamination.
Screening Bacterial Colonies by Hybridization: Intermediate Numbers Protocol Bacterial colonies growing on agar plates are transferred en masse to nitrocellulose filters. The spatial arrangement of colonies on the plates is preserved on the filters. After transfer, the filters are processed for hybridization to an appropriate radiolabeled probe while the original (master) plate is incubated for a few hours to allow the bacterial colonies to regrow in their original positions.
Screening Bacterial Colonies by Hybridization: Small Numbers Protocol Protocol used to screen a small number of bacterial colonies (<200) that are dispersed over several agar plates and are to be screened by hybridization to the same radiolabeled probe. The colonies are gridded onto a master plate and onto a nitrocellulose or nylon filter laid on the surface of a second agar plate.
