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 |
Features:
Large capacity water capture freeze dryer
Tabletop medium-sized freeze dryer
Suitable for drying medium-sized samples
Apply to drying in laboratories such as cosmetics, fermentation broth, soil, materials, etc.
product description:
9kg-12kgs water capture capacity, which can meet the needs of laboratories with large freeze-drying requirements for water capture capacity
-70°C/-90°C ultra-low temperature cold trap, which can capture lower temperature organic solvents
Unique refrigeration technology, the cooling time of the cold trap from the start of the low-temperature refrigeration system to the target value is ≤3 minutes
Direct-capture large coil cold trap, higher efficiency, better ice capture effect
Hemispherical cold trap bottom surface is more conducive to the discharge of condensed water
The cold trap and condenser coil are all Teflon anti-corrosion treatment, which can freeze-dry a large amount of ultra-low temperature organic solvents
One-click hot gas defrost, with defrost compressor protection program.
Large flat flange silicone seal, permanently no need to apply any auxiliary sealing silicone grease
High-precision ultra-wide width detection, vacuum sensor with full-range automatic temperature compensation, making vacuum display more accurate
The cavity vacuum can be read and controlled accurately, and the vacuum control accuracy is ≤0.1mbar
Removable sample shelf with dedicated holes for centrifuge tube placement, making placement and height adjustment easier
Configuration of various types of chambers and T-frames can fully satisfy users’ sample placement
Optional configuration of eutectic point detection system and data export system.
The vacuum pump and sample placement chamber can be combined at will according to user needs, making the configuration more flexible and convenient
Techinical parameter
9-12L Floor tabletop and floor standing one for lab usage
| Tabletop | BD1-9 | BD2-9 | BD1-12 | BD2-12 |
| Floor standing | BH1-9 | BH2-9 | BH1-12 | BH2-12 |
| Water Capture capacity | 9L | 12L | ||
| Cold trap temperature | -70 C | -90℃ | -70C° | -90℃ |
| Cold trap volume | 15L | 20L | ||
| Freeze dried chamber | High permeability organic glass/multi manifold external valve/stainless steel chamber | |||
| Sample shelf | Stainless steel φ 260mm/485mm. 3-10 layers/optional●Electric heating/manual capping/optional | |||
| Vacuum pump | Extreme vacuum 4 * 10-4mbar, pumping capacity ≥ 200L/can be equipped with various types of pumps | |||
| Oil mist filter | Standard configuration | |||
| Vacuum pump connection pipe | Single forming stainless steel flexible pipe DN16 ISO-KF L-1.5m | |||
| anti-corrosive treatment | Cold trap, condensing coil/all equipped with PTFE anti-corrosion treatment as standard. Freeze-drying organic solvents | |||
| External valve | 1-48 ports/optional | |||
| Eutectic point judgment | Support/Optional | |||
| Data audit trail | Support/Optional | |||
| Power | 0.9kw | 1.5kw | 1kw | 1.6KW |
| Size | 390*468*448mm | 480*498*487mm | ||
This kit enables the rapid purification of amplified DNA products from PCR mixes. It is able to effectively remove PCR by-products including primers, dimers, enzymes, unincorporated nucleotides and mineral oil from the desired PCR product. The purified PCR products are fully compatible with restriction enzyme digestion, ligation into vectors, labeling, sequencing and more. This kit can also be used as an alternative to organic extraction and ethanol precipitation to clean up various enzymatic reactions.
The kit is also available in a 96-well format for high-throughput PCR purification. Purification with the 96-well plate can be performed using either a vacuum manifold or centrifugation.
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| Kit Specifications – Spin Column | |
| Column Binding Capacity | 10 μg |
| Size of DNA Purified | 100 – 15,000 bp |
| Average Recovery | > 90% |
| Average Primer Removal | > 90% |
| Minimum Elution Volume | 30 μL |
| Time to Complete 10 Purifications | 15 minutes |
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.
| Component | Cat. 14400 (50 preps) | Cat. 45700 (250 preps) | Cat. 24800 (192 preps) |
|---|---|---|---|
| Binding Buffer C | 30 mL | 5 x 30 mL | 3 x 30 mL |
| Wash Solution A | 12 mL | 2 x 20 mL | 2 x 38 mL |
| Elution Buffer B | 8 mL | 2 x 30 mL | 2 x 15 mL |
| Spin Columns | 50 | 250 | – |
| Collection Tubes | 50 | 250 | – |
| 96-Well Plate | – | – | 2 |
| Adhesive Tape | – | – | 4 |
| 96-Well Collection Plate | – | – | 2 |
| Elution Tubes (1.7 mL) | 50 | 250 | – |
| 96-Well Elution Plate | – | – | 2 |
| Product Insert | 1 | 1 | 2 |