As new technology and methodologies are developed, we will occasionally archive protocols that are no longer commonly used by our partners. These are still available below however.
Leucocyte depletion of 2.0ml of Plasmodium-infected whole blood using MN2100ff cellulose columns
A primary obstacle to whole genome sequencing (WGS) parasite DNA directly from infected whole blood is the overwhelming contamination of human leucocyte DNA (typically >99% of the sample). This challenge can be overcome with methods like hybrid RNA baiting and magnetic separation, however these methods aren’t always amenable to the field. Cellulose powder columns are a low-cost, easy-to-assemble alternative for malaria patient sampling in remote field sites that lack extensive lab facilities.
This protocol involves a leucodepletion cellulose column method that uses MN2100ff as an alternative to Sigma® (Whatman®) CF11 cellulose, which is no longer commercially available. The efficacy of leucodepletion, and ease of blood sampling and filtration was tested in the laboratory and during a pilot field study carried out in 2015. This protocol is provided pre-publication, and is subject to change.