N-(Propargyl-PEG4-carbonyl)-N-bis(PEG1-methyl ester) is a crosslinker that can react with azide compounds or biomolecules via copper catalyzed Click Chemistry to form a stable triazole linkage. The methyl ester groups can be hydrolyzed, reduced, or substituted under different conditions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
N-(Propargyl-PEG4-carbonyl)-N-bis(PEG1-methyl ester) is a crosslinker that can react with azide compounds or biomolecules via copper catalyzed Click Chemistry to form a stable triazole linkage. The methyl ester groups can be hydrolyzed, reduced, or substituted under different conditions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
Sample type purification kit guide
The 16S V2-V3 Library Preparation Kit for Illumina consists of the reagents and components required for library preparation of the 16S V2-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 V2-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 V2-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 V2-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. 70300 (24 preps) | Cat. 70310, 70320, 70330, 70340 (96 preps) |
|---|---|---|---|
| Amplicon PCR (PCR 1) | MGX Master Mix | 330 µL | 1,320 µL |
| 16S V2-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 RNA purification kits isolate total RNA with minimal amounts of genomic DNA contamination. However, for some sensitive downstream applications, it may be desirable to remove all traces of residual DNA. Norgen’s RNase-free DNAse I Kit, with Enzyme Incubation Buffer, can be used for optional on-column DNase digestion with any of Norgen’s RNA purification kits. Alternatively, after isolating total RNA using one of Norgen’s RNA purification kits, the RNA elution can be treated with this DNase I. The RNA can then be purified from the DNase using Norgen’s RNA Clean-Up and Concentration Kit (Cat# 23600), and the RNA can then be used in downstream applications.
Each RNase-Free DNase I Kit is supplied complete with sufficient enzyme and enzyme incubation buffer for 50 or 200 reactions.
Storage Conditions
The DNase I provided is in lyophilized form. It is stable for at least 3 months if stored at room temperature. However, it is recommended to store the DNase I vial at 2 – 8ºC (or below) upon receipt to maintain stability beyond 3 months. Buffer DR and Enzyme Incubation Buffer can be stored at room temperature. After reconstitution with Buffer DR (see product manual), the DNase I should be stored at -20ºC. All reagents should remain stable for at least 1 year in their unopened containers at the appropriate storage temperature.
| Component | Cat. 25710 (50 rxns) | Cat. 25720 (200 rxns) |
|---|---|---|
| DNase I | 1 vial / 1,600 units | 4 vials (1,600 units/vial) |
| Buffer DR | 1 mL | 4 x 1 mL |
| Enzyme Incubation Buffer | 6 mL | 4 x 6 mL |
| Product Insert | 1 | 1 |