Oct 03, 2024
Zymo OneStep PCR Inhibitor Removal Kit
- 1Imperial College London;
- 2Kwame Nkrumah University of Science and Technology;
- 3CMC Vellore
- GEMS - Genomic Environmental Microbial Surveillance
Protocol Citation: Shannon Fitz, Alex Shaw, michael Owusu, Dilip Abraham 2024. Zymo OneStep PCR Inhibitor Removal Kit. protocols.io https://dx.doi.org/10.17504/protocols.io.5qpvokmrbl4o/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: Working
We use this protocol and it's working
Created: October 03, 2024
Last Modified: October 03, 2024
Protocol Integer ID: 108988
Keywords: PCR, PCR inhibitor removal, Purification
Funders Acknowledgement:
The Gates Foundation
Grant ID: INV-049092 PA4273
Abstract
Features:
• For high quality DNA or RNA that is free of enzymatic inhibitors including polyphenolics, humic/fulvic acids, tannins, melanin, etc.
• Fast, one-step procedure for “cleaning” impure samples prior to PCR, sequencing, RT, etc.
Description:
The OneStep‱ PCR Inhibitor Removal Kit contains all the components needed to efficiently remove contaminants from DNA/RNA preparations that can inhibit downstream enzymatic reactions such as PCR and RT. The column matrix has been specifically designed for efficient removal of polyphenolic compounds, humic/fulvic acids, tannins, melanin, etc. from most impure DNA and RNA preparations. Sample cleanup is as simple as: applying, spinning and recovering a sample from the column.
The steps and abstract for this protocol consist entirely of the manufacturer's instructions with no deviations, as per https://files.zymoresearch.com/protocols/_d6030_onestep_pcr_inhibitor_removal_kit.pdf.
Materials
Zymo OneStep PCR Inhibitor Removal KitZymo ResearchCatalog #D6030
Total cost per sample:
£0.5
Approximate time needed per sample:
<10 minutes
Column Preparation
Column Preparation
Insert column into a Collection Tube.
*Please note that matrix in the column may appear dehydrated, or powdery. This is normal.
Open the cap, add 600 µl of Prep-Solution and centrifuge at 8,000 x g for 3 minutes.
Inhibitor Removal:
Transfer the prepared column to a clean 1.5 ml microcentrifuge tube. Add 50 - 200 µl DNA or RNA (in water, TE, or similar) to the Zymo-Spin‱ III-HRC Column and centrifuge at 16,000 x g for 3 minutes.
The filtered DNA (or RNA) is suitable for PCR, (RT), and other downstream applications.
Protocol references
Zymo Research Version 2.0.2-last update, OneStep‱ PCR Inhibitor Removal Kit Cat. No. D6030 (50 spin columns/purifications) Protocol. Available: https://files.zymoresearch.com/protocols/_d6030_onestep_pcr_inhibitor_removal_kit.pdf [2024, October 3].