Both high molecular weight DNA (greater than 1 kb in size; mostly cell associated) and the smaller DNA (150 – 250 bp; derived from the circulation) is effectively isolated and purified using a rapid and convenient spin column protocol. This kit can be used to isolate DNA from a broad range of viruses in urine as well. Salts, metabolic wastes, proteins and other contaminants are removed to yield inhibitor-free DNA for use in sensitive applications. The DNA is of excellent quality for various downstream applications such as PCR, qPCR and DNA fingerprinting, methylation studies and more.
This kit is fully compatible with Norgen’s Urine Collection and Preservation Tubes.
Urine DNA Isolation Kit
This kit provides a fast, reliable and simple procedure for isolating DNA from urine volumes ranging from 50 μL to 1.75 mL of urine. Multiple samples can be processed in 30 minutes.
Urine DNA Isolation Kit (Slurry Format)
This kit provides a fast, reliable and simple procedure for isolating DNA from urine volumes ranging from 3 mL to 25 mL. Multiple samples can be processed in 30 minutes. Multiple samples can be processed in 45 minutes.
Urine DNA Isolation Maxi Kit (Slurry Format)
This kit provides a fast, reliable and simple procedure for isolating DNA from urine volumes ranging from 25 mL of urine up to 80 mL. Multiple samples can be processed in 45 minutes.
Background
DNA found in urine can be divided into 2 basic categories. The larger species, genomic-DNA (gDNA), is generally greater than 1 kb in size, and appears to be derived mainly from exfoliated cells. The second species is smaller, generally between 150 and 250 bp (apoptotic-DNA), and derives, at least in part, from the circulation. The second species is also considered as an RNA/DNA hybrid as reported by Halicka et al. (2000). Both types of DNA can be isolated reliably using this kit.
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| Kit Specifications | |
| Volume of Urine Processed | 1.75 mL |
| Average Yield* | Up to 50 ng |
| Size of DNA Purified | Large (> 1 kb) and small (150-250 bp) |
| Time to Complete 10 Purifications | 30 minutes hands-on time (plus a 1 hour incubation) |
* Yield will vary depending on the type of sample processed
Storage Conditions and Product Stability
All buffers should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment. The kit contains a ready-to-use Proteinase K, which is dissolved in a specially prepared storage buffer. The buffered Proteinase K is stable for up to 2 years after the date of shipment when stored at room temperature.
| Component | Cat. 18100 (50 preps) | Cat. 48800 (50 preps) | Cat. 50100 (50 preps) |
|---|---|---|---|
| Binding Solution K | 15 mL | – | – |
| Slurry B1 | – | 18 mL | 18 mL |
| Proteinase K in Storage Buffer | 2 mL | – | – |
| Pronase K in Storage Buffer | 2 mL | – | – |
| Soluton WN | 9 mL | – | – |
| Binding Buffer A | – | – | 50 mL |
| Lysis Buffer A | – | 30 mL | 30 mL |
| Wash Solution A | – | 38 mL | 38 mL |
| Wash Solution B | 30 mL | – | – |
| Wash Solution D | 9 mL | – | – |
| Binding Solution K | 15 mL | – | – |
| Elution Buffer B | 15 mL | 15 mL | 15 mL |
| Micro Spin Columns | 50 | – | – |
| Mini Filter Spin Columns | – | 50 | 50 |
| Collection Tubes | 50 | 50 | 50 |
| Elution Tubes (1.7 mL) | 100 | 100 | 100 |
| Product Insert | 1 | 1 | 1 |
Norgen’s Fecal Swab Collection and Preservation System is designed for collection, ambient storage and transport of nucleic acids from stool specimens. The Fecal Swab Collection and Preservation System tubes contain Norgen’s Stool Preservative in a liquid format. The user simply collects the specimen using the provided swab, and then transfers the swab into the Stool Preservative. The Stool Preservative prevents the growth of Gram-negative and Gram-positive bacteria and fungi, and also inactivates viruses allowing the resulting non-infectious samples to be handled and shipped safely. In addition, the Stool Preservative eliminates the need to immediately process or freeze samples and allows the samples to be shipped to centralized testing facilities at ambient temperatures. The components of the Stool Preservative allow samples to be stored at room temperature for over 2 years for DNA and 7 days for RNA. To extend the stool RNA stability in the preservative, storage at -20°C or -70°C is recommended upon arrival at the testing facilities until RNA purification.
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| Kit Specifications | |
| Stability of Stool Nucleic Acids at Room Temperature | 2 years for DNA Up to 7 days for RNA* |
* The RNA stability will vary depending on the samples
Storage Conditions and Product Stability
The Fecal Swab Collection and Preservation System (Bulk Format) can be stored at room temperature for up to 2 years without any reduction in kit performance. The expiry date for the swabs is written on the swab wrapper.
| Kit Components | Cat. 45670-B |
|---|---|
| Individually Wrapped Swabs | 50 |
| Fecal Swab Collection and Preservation Tube | 50 |
| Product Insert | 1 |
Sample type purification kit guide
The 16S V3-V4 Library Preparation Kit for Illumina consists of the reagents and components required for library preparation of the 16S V3-V4 amplicon libraries to be used for next-generation sequencing on Illumina platforms. All molecular reagents including primers, enzyme mixes, indexes, and buffers are provided. Instructions for PCR clean up with the AMPure XP Magnetic Beads (supplied by customer) are also included for rapid purification of nucleic acid products generated at two steps of the workflow. The library prep workflow could be used for purified DNA inputs from different sources including stool, soil, water, saliva, plant, urine, skin swab, vaginal swab, cheek swab, nasal swab, plasma/serum, tongue swab, gum swab, and others.
The 16S V3-V4 Library Preparation Kit for Illumina has a streamlined procedure that reduces the handling time such that the library prep procedure can be completed in approximately 4 hours (see diagram below). Input DNA is first subjected to targeted PCR to amplify the V3-V4 region of the DNA encoding 16S rRNA. The post-PCR reaction is then cleaned up using AMPure XP beads. Dual index primers are then added using a limited-cycle PCR. The indexed amplicons flanked by 5′ and 3′ barcoded adaptors are then cleaned using AMPure XP beads. The libraries are then ready for quantification, pooling and sequencing.
| Minimum amount of starting material: | 2.5 µL of DNA (5 ng/µL) |
| Time to complete library preparation: | 4 hours |
Storage Conditions and Product Stability
Norgen’s 16S V3-V4 Library Prep Kit for Illumina is shipped as one kit box (for the 24 prep kit) or two sub-component kits (for the 96 prep kit). All kits should be stored at -20°C upon arrival.
All kit components should remain stable for at least 1 year when stored at the specified storage conditions.
| Step | Component | Cat. 70400 (24 preps) | Cat. 70410, 70420, 70430, 70440 (96 preps) |
|---|---|---|---|
| Amplicon PCR (PCR 1) | MGX Master Mix | 330 µL | 1,320 µL |
| 16S V3-V4 Primer Mix | 70 µL | 280 µL | |
| Index PCR (PCR 2) | Indexing Master Mix | 660 µL | 2 x 1,320 µL |
| N7xx Index Primer | 50 µL | 50 µL | |
| S5xx Index Primer | 70 µL | 70 µL | |
| PCR Clean-Up | Resuspension Buffer | 2 x 1,250 µL | 2 x 5,000 µL |
| Nuclease-free water | 1,250 µL | 1 x 6,000 µL |
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