Water RNA/DNA Purification Kits
For convenient purification of RNA and DNA from microorganisms in water samples
For research use only and NOT intended for in vitro diagnostics.
Powered by Bioz Water RNA/DNA Purification Kits
For convenient purification of RNA and DNA from microorganisms in water samples
Register today to receive an exclusive 15% off* on your first order.
Overview
- Isolate total DNA and RNA from all microorganisms found in water, including bacteria, fungi and algae
- RNA and DNA are both column purified simultaneously using the same column
- Elution contains concentrated DNA and RNA without the need for further precipitation
- Complete RNA (including microRNA) without phenol
- Isolated RNA and DNA are of high quality and integrity for all downstream applications
- Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix
These kits provide a convenient and rapid method for the isolation of microorganisms from water samples. The kits allows for the rapid isolation and purification of total RNA and DNA simultaneously from the microorganisms found in different types of water samples. The total RNA and DNA (including genomic DNA) are isolated from all the microorganisms found in the water, including bacteria, fungi and algae without the use of any inhibitory organic substances The kit purifies genomic DNA, and all sizes of RNA, from large mRNA and ribosomal RNA down to microRNA (miRNA) and small interfering RNA (siRNA). The purified RNA and DNA are highly concentrated, and can be used directly in a number of downstream applications including PCR, qPCR, RT-PCR, qRT-PCR, Northern blotting, Southern blotting and sequencing reactions.
Water RNA/DNA Purification Kit (Spin Column + Filters Kits)
These kits are available with 0.45 μm or 0.22 μm filter formats for capturing microorganisms present in the water.
Water RNA/DNA Purification Kit (Spin Column)
This kit is sold without filters columns.
Details
Supporting Data
Figure 1. High Yield and Purity of RNA and DNA. Total RNA and DNA were simultaneously isolated from 50 mL of water sample containing 107 cfu/mL E.coli using Norgen's Water RNA/DNA Purification Kit and subsequently run on gels for visual analysis. Panel A shows 10 µL aliquots (no RNase treatment) of the 50 µL elutions run on a 1% TAE agarose gel. Genomic DNA and 16S and 23S rRNA bands were visible. Panel B shows 5 µL aliquots (on-column DNase was applied) of the elution run on a 1.5% formaldehyde agarose gel. 16S and 23S rRNA was seen without DNA contamination. From observing the gels it can be seen that the kit allows for the isolation and purification of high yields of concentrated and high quality RNA and DNA.
Figure 2. Detection of E. coli 16S rDNA by Real-Time PCR (SYBR Green) Fifty mL water samples spiked with increasing amounts of E.coli were filtrated through the provided 0.45 µm water filter columns, and RNase-treated DNA was isolated from the filter by following Norgen's Water RNA/DNA Purification Kit manual. Next, 2 µL of DNA from the 50 µL elutions was mixed in 18 µL of total real-time PCR reaction master mix and real-time PCR was performed (95°C for 3 minutes and 40 cycles at 95°C for 15 second and 60°C for 30 seconds.). All tested water samples containing different E.coli amounts were found to correspond to the real-time PCR results. E.coli was specifically detected by E. coli 16S rDNA primers down to 102 cfu/ml, indicating the quality of DNA and the efficiency of filtering system.
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Kit Specifications
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| Minimum Water Input |
10 mL
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| Maximum Water Input |
100 mL
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| Maximum Filter Column Loading Volume |
20 mL
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| Maximum Spin Column Loading Volume |
650 µL
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| Elution Volume |
100 µL
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| Time to Complete 10 Purifications |
45 minutes
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Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years from the date of shipment.
| Component | Cat. 26400 (25 preps) | Cat. 26450 (25 preps) | Cat. 26480 (50 preps) |
|---|---|---|---|
| Lysis Buffer E | 15 mL | 15 mL | 2 x 15 mL |
| Wash Solution A | 18 mL | 18 mL | 38 mL |
| Enzyme Incubation Buffer B | 6 mL | 6 mL | 6 mL |
| Elution Buffer H | 6 mL | 6 mL | 6 mL |
| Mini Spin Columns | 25 | 25 | 50 |
| Filter Columns (0.22 μL) | 25 | - | - |
| Filter Columns (0.45 μL) | - | 25 | - |
| Bead Tubes | 25 | 25 | 50 |
| Collection Tubes | 25 | 25 | 50 |
| Elution Tubes (1.7 mL) | 25 | 25 | 50 |
| Product Insert | 1 | 1 | 1 |
Documentation
FAQs
Spin Column + 0.22 µm Filter, Spin Column + 0.45 µm Filter, Spin Column
Column clogging can result from one or a combination of the following factors:
- Input volume of water sample was too high.
The amount of sample input may need to be decreased to prevent clogging of the Filter Column, particularly for turbid water samples.
- Pre-filtering is necessary.
For highly turbid water samples, or samples containing high levels of sediments, pre-filtration of the sample might be necessary. Pass the sample through a 1-8 µm filter to remove debris prior to applying the sample to the Filter Column.
Lysis was not completed.
- Ensure the filter has not dried out during the 65℃ incubation step. Alternatively, increase the incubation time at 65℃ to 15 minutes.
- Ethanol was not added to the lysate.
Ensure that the appropriate amount of ethanol is added to the lysate before binding to the column.
- Ethanol was not added to the Wash Solution.
Ensure that 42 mL of 96-100% ethanol is added to the supplied Wash Solution A prior to use.
- An alternative elution buffer was used.
It is recommended that the Elution Buffer H supplied with this kit be used for maximum DNA recovery.
If the DNA or RNA does not perform well in downstream applications, it may be due to one or more of the following:
- 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.
- DNA/RNA was not washed twice with the provided Wash Solutions A.
Traces of salt from the binding step may remain in the sample if the column is not washed twice with the Wash Solution A. Salt may interfere with downstream applications and thus must be washed from the column.
- 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
Spin Column with Filters (26400, 26450)
| Title | The Source of the River as a Nursery for Microbial Diversity |
| Journal | PLOS ONE. 2015. |
| Authors | Luiz Felipe Valter de Oliveira, Rogério Margis |
| Title | Assessment of the biological invasion risks associated with a massive outdoor cultivation of the green alga, Pseudochoricystis ellipsoidea |
| Journal | Algal Research. 2015. |
| Authors | Izumi Matsuwaki, Shigeaki Harayama, Misako Kato |
| Title | Characteristics of biofilm development in an operating rainwater storage tank. |
| Journal | Environmental Earth Sciences. 2014. |
| Authors | Kim M, Han M. |
| Title | Amplicon pyrosequencing and ion torrent sequencing of wild duck eubacterial microbiome from fecal samples reveals numerous species linked to human and animal diseases. |
| Journal | F1000 Research. 2013. |
| Authors | Strong T, Dowd S, Gutierrez A, Molnar D, Coffman J. |
| Title | Detection of diverse aquatic microbes in blood and organs of drowning victims: first metagenomic approach using high-throughput 454-pyrosequencing. |
| Journal | Forensic Science International. 2012. |
| Authors | Kakizaki E, Ogura Y, Kozawa S, Nishida S, Uchiyama T, Hayashi T, Yukawa N. |
Spin Column (26480)
| Title | The occurrence patterns of gut bacteria in a post-mined peatland, northern Japan |
| Journal | Mires and Pea. 2022. |
| Authors | Shiro Tsuyuzaki, Tamao Saito, Risa S. Arakawa |
| Title | The Source of the River as a Nursery for Microbial Diversity |
| Journal | PLOS ONE. 2015. |
| Authors | Luiz Felipe Valter de Oliveira, Rogério Margis |
| Title | Assessment of the biological invasion risks associated with a massive outdoor cultivation of the green alga, Pseudochoricystis ellipsoidea |
| Journal | Algal Research. 2015. |
| Authors | Izumi Matsuwaki, Shigeaki Harayama, Misako Kato |
| Title | Characteristics of biofilm development in an operating rainwater storage tank. |
| Journal | Environmental Earth Sciences. 2014. |
| Authors | Kim M, Han M. |
| Title | Amplicon pyrosequencing and ion torrent sequencing of wild duck eubacterial microbiome from fecal samples reveals numerous species linked to human and animal diseases. |
| Journal | F1000 Research. 2013. |
| Authors | Strong T, Dowd S, Gutierrez A, Molnar D, Coffman J. |
| Title | Detection of diverse aquatic microbes in blood and organs of drowning victims: first metagenomic approach using high-throughput 454-pyrosequencing. |
| Journal | Forensic Science International. 2012. |
| Authors | Kakizaki E, Ogura Y, Kozawa S, Nishida S, Uchiyama T, Hayashi T, Yukawa N. |
