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INSTRUCTION MANUAL
ZymoBIOMICS
Catalog Nos. D4300T, D4300, & D4304
Highlights
•
Validated Unbiased for Microbiome Measurements: Unbiased cellular lysis validated using the
ZymoBIOMICS Microbial Community Standard.
•
Inhibitor-Free DNA from Any Sample: Isolate ultra-pure DNA ready for any downstream application.
•
Certified Low Bioburden: Boost your detection limit for low abundance microbes.
•
Simple Workflow: Simply bead-beat sample, purify via spin-column, and filter to remove PCR inhibitors.
No precipitation or lengthy incubations!
For Research Use Only
™
DNA Miniprep Kit
Contents
Product Contents & Specifications ................................. 1
Product Description ..................................................... 2-3
Protocol ........................................................................ 4-5
Appendices
A. Sample Collection ............................................... 6
B. Application Notes ........................................... 7-10
ZymoBIOMICS
D. Troubleshooting ........................................... 12-13
Ordering Information ..................................................... 14
™
Microbial Standards ....... 11-12
Ver. 1.4.1
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Summary of Contents for Zymo Research ZymoBIOMICS D4300T
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Page 1: Table Of Contents
INSTRUCTION MANUAL ZymoBIOMICS ™ DNA Miniprep Kit Catalog Nos. D4300T, D4300, & D4304 Highlights • Validated Unbiased for Microbiome Measurements: Unbiased cellular lysis validated using the ZymoBIOMICS Microbial Community Standard. • Inhibitor-Free DNA from Any Sample: Isolate ultra-pure DNA ready for any downstream application. •... -
Page 2: Product Contents & Specifications
• ® Equipment – Microcentrifuge, vortex/Disruptor Genie high speed cell disrupter (recommended). organisms including infectious agents (see Appendix A). ZYMO RESEARCH CORP. Phone: (949) 679-1190 ▪ Toll Free: (888) 882-9682 ▪ Fax: (949) 266-9452 ▪ info@zymoresearch.com ▪ www.zymoresearch.com... -
Page 3: Product Description
, as shown in Figure 3. In addition, the representation of organisms ™ ZymoBIOMICS™ DNA Miniprep Kit is equipped with Zymo Research’s proprietary OneStep PCR Inhibitor in downstream metagenomic analyses that is not Removal technology enabling PCR from the most PCR prohibitive environmental samples rich in humic and reflective of actual fulvic acids, tannins, melanin, and other polyphenolic compounds. - Page 4 (2x250bp). Overlapping paired-end reads were assembled into complete amplicon sequences. Amplicon sequences were profiled with Qiime using Greengenes 16S rRNA gene database (gg_13_8). ZYMO RESEARCH CORP. Phone: (949) 679-1190 ▪ Toll Free: (888) 882-9682 ▪ Fax: (949) 266-9452 ▪ info@zymoresearch.com ▪ www.zymoresearch.com...
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Page 5: Protocol
DNA Wash Buffer 2 to the Zymo-Spin ™ IICR Column in a Collection Tube and centrifuge at 10,000 x g for 1 minute. Discard the flow-through. ZYMO RESEARCH CORP. Phone: (949) 679-1190 ▪ Toll Free: (888) 882-9682 ▪ Fax: (949) 266-9452 ▪ info@zymoresearch.com ▪ www.zymoresearch.com... - Page 6 1.5 ml microcentrifuge tube and centrifuge at exactly 16,000 x g for 3 minutes. The filtered DNA is now suitable for PCR and other downstream applications. ZYMO RESEARCH CORP. Phone: (949) 679-1190 ▪ Toll Free: (888) 882-9682 ▪ Fax: (949) 266-9452 ▪ info@zymoresearch.com ▪ www.zymoresearch.com...
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Page 7: Sample Collection
Poole and Prof. A. Mehle, Department of Medical Microbiology and Immunology, University of Wisconsin, Madison; Ebola (Kikwit) - L. Avena and Dr. A. Griffiths, Department of Virology and Immunology, Texas Biomedical Research Institute; HSV-1/2. ZYMO RESEARCH CORP. Phone: (949) 679-1190 ▪ Toll Free: (888) 882-9682 ▪ Fax: (949) 266-9452 ▪ info@zymoresearch.com ▪ www.zymoresearch.com... -
Page 8: Application Notes
3. Cut the filter and place directly into the ZR BashingBead Lysis Tubes (0.1 & 0.5 mm) and proceed to Step 1 (page 4). ZYMO RESEARCH CORP. Phone: (949) 679-1190 ▪ Toll Free: (888) 882-9682 ▪ Fax: (949) 266-9452 ▪ info@zymoresearch.com ▪ www.zymoresearch.com... - Page 9 Lysis Tube (0.1 & 0.5 mm) and then add 750 µl ZymoBIOMICS™ Lysis Solution. 4. Continue to Step 2 (page 4) and proceed with the remaining protocol as written. ZYMO RESEARCH CORP. Phone: (949) 679-1190 ▪ Toll Free: (888) 882-9682 ▪ Fax: (949) 266-9452 ▪ info@zymoresearch.com ▪ www.zymoresearch.com...
- Page 10 (6.5 m/s) for 5 minutes. The extracted DNA was then subjected to microbial composition profiling via 16S rRNA gene targeted sequencing. Experiment was performed in technical duplicates ZYMO RESEARCH CORP. Phone: (949) 679-1190 ▪ Toll Free: (888) 882-9682 ▪ Fax: (949) 266-9452 ▪ info@zymoresearch.com ▪ www.zymoresearch.com...
- Page 11 16S rRNA gene targeted sequencing. Experiment was performed in technical duplicates. 0 Day 2 Weeks 1 Month ZYMO RESEARCH CORP. Phone: (949) 679-1190 ▪ Toll Free: (888) 882-9682 ▪ Fax: (949) 266-9452 ▪ info@zymoresearch.com ▪ www.zymoresearch.com...
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Page 12: Standardize Sample Preparation With
It is ideal for assessing bias of DNA extraction methods since it contains three easy-to-lyse Gram-negative bacteria (e.g. Escherichia coli), five tough-to-lyse Gram-positive bacteria (e.g. Listeria monocytogenes), and two tough-to-lyse yeasts (e.g. Saccharomyces cerevisiae). Bead Beating Device Calibration Protocol: Zymo Research suggests calibrating bead beating devices with the ZymoBIOMICS ™ Microbial Community Standard in order to ensure bias â... -
Page 13: Troubleshooting
Make sure bags of columns and buffer bottles are properly sealed for storage. Use of these outside a clean room or hood can result in contamination. ZYMO RESEARCH CORP. Phone: (949) 679-1190 ▪ Toll Free: (888) 882-9682 ▪ Fax: (949) 266-9452 ▪ info@zymoresearch.com ▪ www.zymoresearch.com... - Page 14 3 minutes, and centrifuge again to increase yield without further dilution. ZYMO RESEARCH CORP. Phone: (949) 679-1190 ▪ Toll Free: (888) 882-9682 ▪ Fax: (949) 266-9452 ▪ info@zymoresearch.com ▪ www.zymoresearch.com...
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Page 15: Ordering Information
250 ml ™ ™ ™ ZymoBIOMICS , Zymo-Spin , and DNA/RNA Shield are trademarks of Zymo Research Corp. Disruptor Genie ® is a registered trademark of Scientific Industries, Inc. FastPrep ® is a registered trademark of Qbiogene, Inc. Illumnia ®...
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