This kit is designed for the rapid spin column preparation of genomic DNA from 2 x 109 viable bacterial cells (between 0.5 and 1.0 mL of culture). This kit can be used for both Gram-negative and Gram-positive bacteria including Escherichia coli and Bacillus cereus. Purified genomic DNA is of an excellent quality and yield, and is fully compatible with restriction enzyme digestions, sequencing, PCR, qPCR and more. Also available in 96-well format for high throughput applications.
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| Kit Specifications (96-well) | |
| Maximum Input | 2 x 109 bacterial cells |
| Well Binding Capacity | 25 µg |
| Average Yield* | Up to 20 µg* |
| Time to Complete 10 Purifications | 90 minutes |
* Yield will vary depending on the type of sample processed
Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 1 year 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 1 year after the date of shipment when stored at room temperature.
| Component | Cat. 17900 (50 preps) | Cat. 17950 (192 preps) |
|---|---|---|
| Resuspension Solution A | 20 mL | 60 mL |
| Lysis Buffer P | 18 mL | 60 mL |
| Solution BX | 28 mL | 110 mL |
| Wash Solution A | 18 mL | 2 x 38 mL |
| Elution Buffer B | 30 mL | 2 x 30 mL |
| Proteinase K | 12 mg | 50 mg |
| Spin Columns | 50 | – |
| 96-Well Plate | – | 2 |
| Adhesive Tape | – | 4 |
| Collection Plate | – | 2 |
| Collection Tubes | 50 | – |
| Elution Tubes (1.7 mL) | 50 | – |
| Elution Plate | – | 2 |
| Product Insert | 1 | 1 |
Sample type purification kit guide
The 16S V1-V3 Library Preparation Kit for Illumina consists of the reagents and components required for library preparation of the 16S V1-V3 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 V1-V3 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 V1-V3 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.
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| 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 V1-V3 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. 70200 (24 preps) | Cat. 70210, 70220, 70230, 70240 (96 preps) |
|---|---|---|---|
| Amplicon PCR (PCR 1) | MGX Master Mix | 330 µL | 1,320 µL |
| 16S V1-V3 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 |
Norgen’s Cells and Tissue DNA Isolation Kits are designed for the rapid preparation of genomic DNA from cultured cells as well as various tissue samples and urine. The purified genomic DNA is fully digestible with all restriction enzymes tested, and is completely compatible with PCR and Southern Blot analysis.
Cells and Tissue DNA Isolation Kit (Spin Column)
Purification is based on spin column chromatography as the separation matrix. Norgen’s columns bind DNA under optimized salt concentrations and release the bound DNA under low salt and slightly alkali conditions. The protocol can be completed in approximately 30 minutes for cells and within 90 minutes for tissues. Each kit contains sufficient materials for 50 preparations.
Cells and Tissue DNA Isolation Micro Kit (Micro)
Optimized for small inputs of cells and tissues, such as Laser-Captured Microdissection (LCM). Purification is based on spin column chromatography as the separation matrix. Norgen’s columns bind DNA under optimized salt concentrations and release the bound DNA under low salt and slightly alkali conditions. Preparation time for a single sample is approximately 60 minutes, and each kit contains sufficient materials for 50 preparations.
Cells and Tissue DNA Isolation Kit (Magnetic Bead System)
Purification is based on the use of magnetic beads that bind DNA under optimized binding conditions. Norgen’s Cells and Tissue DNA Isolation Kit (Magnetic Bead System) allows for the isolation of genomic DNA from various types of animal tissues or cell samples. Preparation for 10 purifications is approximately 40 minutes of hands-on time.
Cells and Tissue DNA Isolation 96-Well Kit (High Throughput Magnetic Bead System)
Purification is based on the use of magnetic beads that bind DNA under optimized binding conditions. Norgen’s Cells and Tissue DNA Isolation 96-Well Kit (Magnetic Bead System) allows for the isolation of genomic DNA from various types of animal tissues or cell samples. The Cells and Tissue DNA Isolation 96-Well Kit (Magnetic Bead System) also can be integrated with a robotic automation system.
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| Kit Specifications | |
| Maximum Input | 20 mg of animal tissue Up to 150 μL of viral suspension 3 x 106 cells |
| Column Binding Capacity | > 50 μg |
| Average Yield | 8 μg (from 1 x 106 HeLa Cells) 10 μg (from 10 mg kidney) |
| Elution Volume | 50 – 200 μL |
| Analyte Purified | Genomic DNA, mitochondrial DNA, viral DNA |
| Format | Spin Column |
| Time to Complete 10 Purifications | 30 min (cells) and 90 min (tissue) |
Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 1 year 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 1 year after the date of shipment when stored at room temperature.
| Component | Cat. 53100 (50 preps) | Cat. 57300 (50 preps) | Cat. 59100 (50 preps) | Cat. 62500 (192 preps) |
|---|---|---|---|---|
| Lysis Buffer B | 20 mL | 20 mL | 20 mL | 1 x 40 mL 1 x 20 mL |
| Solution WN | 18 mL | 18 mL | 18 mL | 55 mL |
| Wash Solution A | 18 mL | 18 mL | – | – |
| Elution Buffer B | 30 mL | 8 mL | 15 mL | 1 x 30 mL 1 x 15 mL |
| Proteinase K | 1.2 mL | 1.2 mL | 1.2 mL | – |
| Proteinase K in Sotrage Buffer | – | – | – | 4 mL |
| Magnetic Bead Suspension | – | – | 2 x 1.1 mL | 8.5 mL |
| 96-Well Plate | – | – | – | 2 |
| Spin Columns | 50 | – | – | – |
| Micro Spin Columns | – | 50 | – | – |
| Collection Tubes | 50 | 50 | – | – |
| Elution Tubes (1.7 mL) | 50 | 50 | 50 | – |
| 96-Well Elution Plate | – | – | – | 2 |
| Adhesive Tape | – | – | – | 2 |
| Product Insert | 1 | 1 | 1 | 1 |