Aug 15, 2025
Version 1
miRNA extraction from fecal matter V.1
- 1Colorado State University
- NHeyer
Protocol Citation: nick Heyer 2025. miRNA extraction from fecal matter. protocols.io https://dx.doi.org/10.17504/protocols.io.kxygx43pdl8j/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: In development
We are still developing and optimizing this protocol
Created: August 14, 2025
Last Modified: August 15, 2025
Protocol Integer ID: 224732
Keywords: mirna extraction from fecal matter, mirna extraction for tissue, mirna extraction, qiagen proticol, modification to the qiagen proticol, fecal matter, tissue
Disclaimer
This Proticol is still being eddited
Abstract
This is a modification to the Qiagen proticol for miRNA extraction for tissues
Prep
Set Centrifuge to 4C if needed
Create 70% and 90% Et-OH
______num samples
______num samples
______num samples
______num samples
Lysis, chloroform extraction, and Large Nucliotide removal
27m
In a 1.5ml tube, homogenize _________mg of dry fecal matter in 700ul of QIAzol
10m
Incubate at 20C for 5min
Add 140ul of chloroform, MIX WELL and let incubate for 1 min at 20C, MIX AGAIN, and incubate for 3 min
Centrifuge at >12,000xg at 4C for 15 min
15m
Transfer aqueous phase and determine volume by pipetting, combine with 1x Volume of 70% Et-OH .
2m
Run through a "RNeasy Mini spin Column" at >8,000xg at 20C
miRNA recovery
Recover flow through, and add 0.65x volumes for 100% Et-OH
Run through a "RNeasy Mini spin Column" at >8,000xg at 20C, 700ul at a time discarding flow-through
Wash matrix with 700ul RWT Buffer
Wash matrix with 500 ul of RPE
Wash matrix ______x with ice cold 90% Et-OH
Dry Tube by spinning column for 5 min
Elute in 30ul of elution buffer