Training for stool-based qPCR diagnostics for soil-transmitted helminths developed by London Centre for Neglected Tropical Disease Research

This video demonstrates a tutorial on running a script written in Python (provided by Panos Bozelos), for compiling the individually exported qPCR file outputs, as ‘.xlsx’ files (demonstrated in CLIP4) into a single master file for downstream analysis in R or equivalent.

This video shows: i) the qPCR software set-up for the detection of soil-transmitted helminths, and ii) the review of previous qPCR runs with some troubleshooting. We have used the StepOne Plus instrument from Thermofisher for the demonstration of “Standard curve/Absolute quantitation”, advanced set up. Visit your instrument’s manual for more details and guidance with the set-up for your work.

This video demonstrates: i) the serial dilutions of DNA standards (or positive controls), ii) the loading of DNA samples in the qPCR, 96-well plate, and iii) the sealing and spinning of the qPCR plate. The 96-well template introduced in this video covers the screening of 39 DNA samples in duplicate, DNA standards in triplicate and non-template controls, also in triplicate. Each plate contains the screening of a single target (e.g., single STH species or extraction control). The video contains tips and explanations at each step of the process (Note: instructions are not a substitute for following standard operating procedures). Total duration 14 minutes

This video demonstrates: i) the resuspension of primers/probes used in qPCR testing, ii) the preparation of the master mix for qPCR reactions and the 96-well plate loading, and, iii) the aliquoting of the reagent mix (a mix containing enzyme, salts etc). The video contains tips and explanations at each step of the process (Note: instructions are not a substitute for following standard operating procedures). Total duration 9 minutes

This video demonstrates the extraction of DNA from stool samples, using the FAST SPIN kit for soil from MP Biomedicals. The video contains tips and explanations at each step of the process (Note: instructions are not a substitute for following standard operating procedures). Total duration 42 minutes