Sample type purification kit guide
The 16S V1-V2 Library Preparation Kit for Illumina consists of the reagents and components required for library preparation of the 16S V1-V2 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-V2 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-V2 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-V2 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. 70100 (24 preps) | Cat. 70110, 70120, 70130, 70140 (96 preps) |
|---|---|---|---|
| Amplicon PCR (PCR 1) | MGX Master Mix | 330 µL | 1,320 µL |
| 16S V1-V2 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 |
TCO-PEG2-DBCO is a short PEG linker featuring a trans-cyclooctene and a DBCO group. DBCO is a click chemistry handle which easily reacts with azides, while the TCO function readily reacts with tetrazine-containing compounds.
TCO-PEG2-DBCO is a short PEG linker featuring a trans-cyclooctene and a DBCO group. DBCO is a click chemistry handle which easily reacts with azides, while the TCO function readily reacts with tetrazine-containing compounds.
RNA/DNA/Protein Purification Plus Kit
This kit provides a rapid method for the high throughput isolation and purification of total RNA, DNA and proteins sequentially from a single sample of cultured animal cells, small tissue samples, blood, bacteria, or yeast. The kit employs two columns: 1) for gDNA purification and 2) for RNA purification utilizing Norgen’s resin columns (superior for the binding of all RNA sizes including miRNA). The proteins are also purified on the second column after RNA elution. The proteins are eluted in buffer and are ready for downstream application without any further clean up required. The proteins can be quantified directly, used in western blots, ELISA or mass spectrometry. This kit provides a rapid spin-column method for the isolation and purification of total RNA, genomic DNA and proteins sequentially from a single sample of cultured animal cells, small tissue samples, blood, bacteria, yeast, fungi or plants.
RNA/DNA/Protein Purification Plus Micro Kit
This kit provides a rapid spin-column method for the isolation and purification of total RNA, DNA and proteins sequentially from a single sample of cultured animal cells, small tissue samples, microdissected samples including LCM, stem cells, sorted cells, and CTC. The total RNA, genomic DNA and proteins are all column purified in less than 30 minutes. The RNA and DNA can be eluted in as little as 20 µL while the protein can be eluted in as little as 50 µL. This kit provides the same performance as if the samples were isolated from dedicated kits.
RNA/DNA/Protein Purification 96-Well Plus Kit
The kit employs two plates: 1) for DNA purification and 2) for RNA purification utilizing Norgen’s resin (superior for the binding of all RNA sizes including miRNA). Please see the protocol schematic below.
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| Kit Specifications | |
| Maximum Column Binding Capacity | 35 μg for RNA 10 μg for DNA 100 μg for protein |
| Maximum Column Loading Volume | 650 μL |
| Size of RNA Purified | All sizes, including small RNA (< 200 nt) |
| Size of DNA Purified | ≥ 30 kb |
| Time to Complete 10 Purifications | 30 minutes |
| Maximum Amount of Starting Material:Animal CellsAnimal TissuesBloodPlant / Fungi Tissues | 5 x 105 cells5 mg (for selected tissues)50 µL25 mg |
| Average Yield* HeLa Cells (5 x 105 cells) HeLa Cells (5 x 105 cells) HeLa Cells (5 x 105 cells) | RNA: 7.5 μg DNA: 1-2 μg Protein: 70 μg |
Storage Conditions and Product Stability
The Protein Loading Dye should be stored at -20°C after the addition of DL-Dithiothreitol (DTT). All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 1 year after the date of shipment.
| Component | Cat. 47700 (50 preps) | Cat. 51600 (50 preps) | Cat. 51700 (96 preps) |
|---|---|---|---|
| Buffer SKP | 40 mL | 40 mL | – |
| Lysis Buffer Q | – | – | 40 mL |
| Wash Solution A | 2 x 38 mL 1 x 18 mL | 2 x 38 mL 1 x 18 mL | 2 x 38 mL 1 x 18 mL |
| Elution Solution A | 6 mL | 6 mL | 20 mL |
| Elution Buffer F | 15 mL | 15 mL | 2 x 15 mL |
| Wash Solution C | 30 mL | 30 mL | 60 mL |
| Binding Buffer A | 8 mL | 8 mL | 8 mL |
| Elution Buffer C | 8 mL | 8 mL | 30 mL |
| Protein Neutralizer | 4 mL | 4 mL | 4 mL |
| Protein Loading Dye | 2 mL | 2 mL | 3 x 2 mL |
| gDNA Purification Columns | 50 | – | – |
| gDNA Purification Micro Columns | – | 50 | – |
| gDNA Purification 96-Well Plate | – | – | 1 |
| RNA/Protein Purification Columns | 50 | – | – |
| RNA/Protein Purification Micro Columns | – | 50 | – |
| RNA/Protein Purification 96-Well Plate | – | – | 1 |
| Collection Tubes | 150 | 150 | – |
| Collection Plate | – | – | 5 |
| Elution Tubes (1.7 mL) | 150 | 150 | – |
| Elution Plate | – | – | 3 |
| Lysis Preparation Plate | – | – | 2 |
| Adhesive Tape | – | – | 4 |
| Product Insert | 1 | 1 | 1 |