Microbiome DNA Isolation Kit

Overview

Universal method to isolate and detect microorganisms and host cells simultaneously
For use with samples collected using a swab - fresh, frozen or preserved in Norgen's Swab Collection and DNA Preservation System or Norgen's Fecal Swab Collection and Preservation System
Rapid and convenient spin-column format
Remove all PCR inhibitors from DNA samples
Isolate high quality total DNA for PCR and NGS applications (Microbiome/Metagenomic sequencing)
Optimized to isolate from the full preservative volume of the Swab Collection and DNA Preservation System for maximum yield

Norgen's Microbiome DNA Isolation Kit provides a convenient and rapid method to isolate total DNA from a variety of Microbiome samples collected using a swab. This kit can be used for fresh (dry transport) swab or any swab preserved using Norgen's Swab Collection and DNA Preservation Kit or any other of Norgen's Preservation Systems. This universal protocol conveniently allows for the isolation of total genomic DNA from both host and microbial cells found in the swab sample simultaneously. The kit removes all traces of PCR impurities using the provided combination of chemical and physical homogenization and lysis. A simple and rapid spin column procedure is then used to further purify the DNA. The purified DNA is of the highest quality and is fully compatible with downstream PCR or metagenomics using NGS.

Details

Kit Specifications
Maximum sample input	1 mL of preserved swab sample* or up to 0.5 mL preserved samples**
Swab samples tested	Fecal, saliva, buccal, food, nasal, blood, surface, skin
Maximum Column Binding Capacity	50 μg
Maximum Column Loading Volume	650 μL
Time to Complete 10 Purifications	30 minutes

* Collected using Norgen’s Swab Collection and DNA Preservation Kit (Cat. 45690)
**Please see the table in Section B of the Product Insert - Samples Collected using Norgen's Preservation Devices

Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 1 year from the date of shipment.

Component	Cat. 64100 (50 preps)
Lysis Buffer E	55 mL
Lysis Additive A	6 mL
Binding Buffer I	7 mL
Binding Buffer B	30 mL
Wash Solution A	18 mL
Elution Buffer B	8 mL
Mini Spin Columns	50
Collection Tubes	50
Elution Tubes (1.7 mL)	50
Product Insert	1

Documentation

PROTOCOLS

(64100) Microbiome DNA Isolation Kit - Protocol (50 preps)

SAFETY DATA SHEETS

64100 - Microbiome DNA Isolation Kit - SDS

FAQs

Spin Column

Why do I have poor DNA recovery?

Poor DNA recovery could be due to one or more of the following:

An alternative elution buffer was used.
It is recommended that the Elution Buffer B supplied with this kit be used for maximum DNA recovery.
Lysis Additive A was not added to the lysate.
Ensure that the provided Lysis Additive A is added to increase DNA yield. Also, an incubation can be performed at 65°C for 10 minutes after addition of the Lysis Additive A and prior to vortexing to maximize DNA recovery.
Ethanol was not added to the lysate.
Ensure that an equal amount of ethanol is added to the lysate before binding to the column.
Ethanol was not added to the Wash Solution A.
Ensure that 42 mL of 96 - 100% ethanol is added to the supplied Wash Solution A prior to use.

Why is the purified DNA not performing well in downstream applications?

If the DNA does not perform well in downstream applications, it may be due to one or more of the following:

Elution is brown.
Ensure that the Lysis Additive A is added. Also ensure Binding Solution I is added to the lysate and that it is incubated on ice for 10 minutes prior to spinning down the lysate. Avoid any contact with the pellet or surface residue when collecting the supernatant after the 5-minute spin during Sample Preparation.
Lysis Additive A was not added to the lysate.
Ensure that the provided Lysis Additive A is added to the lysate.
DNA was not washed with the provided Binding Buffer B and Wash Solution A.
Traces of salt from the binding step may remain in the sample if the column is not washed three times with the provided Binding Buffer B and Wash Solution A. Salt may interfere with downstream applications, and thus must be washed from the column.
Ethanol carryover.
Ensure that the dry spin under the Column Wash procedure is performed in order to remove traces of ethanol prior to elution. Ethanol is known to interfere with many downstream applications.
Binding Buffer I was not added to the lysate.
Ensure that the Binding Buffer I is added to the lysate and that it is incubated on ice for 10 minutes prior to spinning down the lysate.
PCR reaction conditions need to be optimized.
Take steps to optimize the PCR conditions being used, including varying the amount of template, changing the source of Taq polymerase, looking into the primer design, and adjusting the annealing conditions.

Citations

Title	Transcript and blood-microbiome analysis towards a blood diagnostic tool for goats affected by Haemonchus contortus
Journal	Scientific Reports. 2022.
Authors	Yonathan Tilahun, Jessica Quijada Pinango, Felicia Johnson, Charles Lett, Kayla Smith, Terry Gipson, Malcolm McCallum, Peter Hoyt, Andrew Tritt, Archana Yadav, Mostafa Elshahed & Zaisen Wang

Title	First line of defence: Skin microbiota may protect anurans from infective larval lungworms
Journal	International Journal for Parasitology: Parasites and Wildlife. 2021.
Authors	Keit Christian, Richard Shine, Kimberley A. Day, Mirjam Kaestli, Karen Gibb, Catherine M. Shilton, Gregory P. Brown