OPTIMISATION OF WILD BIRDS’ FECAL EXTRACTION USING MACHEREY-NAGEL NUCLEOSPIN DNA STOOL KIT

Current DNA extraction methods for bird faeces samples might not provide sufficient DNA quantity, which in turn affects downstream analyses. This study aimed to improve the current method of DNA extraction from the faeces of wild birds - a total of seven (n = 7) samples for analysis and protocol comparison. Three different methods in this study were compared: (1) The original method as per the manufacturer’s instructions, (2) a supplementary chicken-specific protocol provided by the manufacturer, and (3) modifications to the protocol conducted in the laboratory. After extensive troubleshooting, DNA yield was significantly increased by utilising the modified protocol, which includes ethanol evaporation with a heat block, a direct 30-minutes freezer step during contaminant precipitation replacing ice, and a warm elution buffer for improved DNA elution. These findings offer valuable guidance for developing a refined and more specific methodology for collecting and processing bird faeces samples, improving the reliability of DNA-based studies in microbiota research in the bird gut.