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September 23, 2025Open Access

Modified Three Peaks protocol - Power Soil DNA Isolation Kit v1

Key Points

The protocol achieves a median yield of ~40 mg of total dna per sample, facilitating multiple dna-based analyses.
Three distinct lysis steps optimize microbial lysis and ensure effective dna extraction from samples.
High-speed centrifugation is used between lysis steps to separate unlysed cells from the dna-containing supernatant.
This enhanced protocol allows for improved outcomes in sequencing and other dna applications, highlighting its utility.

Abstract

DNA extraction protocol optimized to maximize DNA yield, fragment length and microbial lysis. This protocol is a modified version of https://www.protocols.io/view/the-39-three-peaks-39-faecal-dna-extraction-method-kqdg34m9pl25/v2 The Three Peaks protocol is characterized by three separate lysis steps: (i) a 10% SDS-based lysis, (ii) a ProteinaseK-based lysis, (iii) a bead-beating-based lysis (optimized power and time settings). In between lysis steps, high-speed centrifugation collects the unlysed cell pellet at the bottom, leaving the supernatant containing DNA to be collected in a separate tube. At the end of the lysis steps three separate tubes containing DNA in a supernatant are processed separately with a column-based DNA isolation procedure, using the Power Soil Pro Kit (QIAGEN). The protocol yields ~40 mg (median) of total DNA/sample, ensuring enough DNA for multiple sequencing and other DNA-based procedures.

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