N-(Boc-PEG3)-N-bis-(PEG3-Amino-Tri-(Propargyl-PEG2-ethoxymethyl)-methane) is a crosslinker consisting of six propargyl groups and a t-Boc protected aminooxy group. The propargyl groups can form triazole linkage with azide-bearing compounds or biomolecules via copper catalyzed Click Chemistry. The protected amine can be deprotected under acidic conditions. The hydrophilic PEG spacer increases solubility in aqueous media. Reagent grade, for research use only.
N-(Boc-PEG3)-N-bis-(PEG3-Amino-Tri-(Propargyl-PEG2-ethoxymethyl)-methane) is a crosslinker consisting of six propargyl groups and a t-Boc protected aminooxy group. The propargyl groups can form triazole linkage with azide-bearing compounds or biomolecules via copper catalyzed Click Chemistry. The protected amine can be deprotected under acidic conditions. The hydrophilic PEG spacer increases solubility in aqueous media. Reagent grade, for research use only.
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 |
The MagPure Plasmid purification system uses the paramagnetic bead technology for high-throughput preparation of high-copy or low-copy plasmid DNA from E. coli cells. This kit also can be used with fosmid and BAC vector-based constructs. The system uses alkaline lysis followed by a MagPure purification to differentially bind plasmid DNA to paramagnetic beads. While the DNA is bound to the beads, contaminants can be rinsed away using a simple washing procedure. Because MagPure uses magnetic separation technology, the protocol does not require vacuum filtration. This makes kit extremely amenable to automation. Plasmid DNA purified with this system is most commonly used in Sanger Sequencing and PCR amplification.
Specifications
| Features | Specifications |
| Main Functions | Isolation up to 20µg plasmid DNA from 1-3ml bacterial culture |
| Applications | Enzyme digestion, sequencing, PCR and labeling, etc. |
| Purification technology | Magnetic beads technology |
| Process method | Manual or automatic |
| Sample type | Conventional plasmid, plasmid≤30KB |
| Sample amount | 1-3ml |
| Elution volume | ≥50μl |
This product is based on the purification method of high binding magnetic particles. The sample is lysed and digested under the action of lysate and Lysozyme. DNA is released into the lysate. After adding magnetic particles and binding solution, DNA will be adsorbed on the surface of magnetic particles, and impurities such as proteins will be removed without adsorption.The adsorbed particles were washed with washing solution to remove proteins and impurities, washed with ethanol to remove salts, and finally DNA was eluted by Elution Buffer.
Advantages
1. Suitable for extracting plasmids from 1-5ml or <3ml YT medium.
2. The same amount of buffer 1, 2, and 3 avoids errors caused by adjusting the pipette, making it convenient to use in conjunction with automated workstations.
3. Containing buffer 1 for washing, reducing the problem of false high production.
4. The purified plasmid can be directly used for sequencing, enzyme digestion, PCR, and other applications.
Kit Contents
| Contents | P181102 | P181103 | P181104 |
| Purification Times | 100 Preps | 500 Preps | 5000 Preps |
| RNase A | 10 mg | 50 mg | 2 x 250 mg |
| Buffer P1 | 30 ml | 150 ml | 2 x 750 ml |
| Buffer P2 | 30 ml | 150 ml | 2 x 750 ml |
| Buffer N3 | 30 ml | 150 ml | 2 x 750 ml |
| Buffer PW1 | 35 ml | 180 ml | 2 x 900 ml |
| MagPure Particle NB* | 2.2 ml | 11 ml | 2 x 60 ml |
Storage and Stability
RNase A and MagPure Particle NB should be stored at 2-8°C upon arrival. However, short-term storage (up to 24 weeks) at room temperature (15-25°C) does not affect its performance. The remaining kit components can be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions. After addition of RNase A, Buffer P1 is stable for 6 months when stored at 2-8°C.
Experiment Data
The MagPure Plasmid purification system uses the paramagnetic bead technology for high-throughput preparation of high-copy or low-copy plasmid DNA from E. coli cells. This kit also can be used with fosmid and BAC vector-based constructs. The system uses alkaline lysis followed by a MagPure purification to differentially bind plasmid DNA to paramagnetic beads. While the DNA is bound to the beads, contaminants can be rinsed away using a simple washing procedure. Because MagPure uses magnetic separation technology, the protocol does not require vacuum filtration. This makes kit extremely amenable to automation. Plasmid DNA purified with this system is most commonly used in Sanger Sequencing and PCR amplification.
