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 |
Nucleic acid testing (NAT) is the method of choice for detection and quantification of a wide range of micro organisms. Primerdesign manufactures and supplies high quality quantitative real-time PCR kits for the detection of numerous significant pathogens. The genesigPLEX range provides the most efficient method of detecting DNA and RNA by combining multiple assays into a single tube. This principle of multiplexing represents state of the art PCR testing and is a key feature of the high-throughput testing conducted by laboratories around the world. These assays include process controls to verify the quality of the nucleic acid extraction and eliminate false negative results.
The kit is designed with the broadest possible detection profile to ensure that all clinically relevant strains and subtypes are detected. Target sequences are selected by working with data from key opinion leaders in the field. Multiple sequence alignments and unprecedented real-time PCR expertise in design and validation ensure the best possible kit.
Details of the target and priming specificity are included in the handbook.
Packaged, optimised and ready to use. Expect Better Data.
Precise reproducible results
Time saving through completing multiple tests in one go
Cost saving through a reduced price compared with buying four kits separately
Exceptional value for money
Supplied lyophilised – no cold chain shipping
Bis-propargyl-PEG14 is a homobifunctional crosslinker that can participate in Click Chemistry to yield a stable triazole linkage with azide; copper is needed as a catalyst. With PEG chain embedded in the molecule, the hydrophilicity is greatly improved. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
Bis-propargyl-PEG14 is a homobifunctional crosslinker that can participate in Click Chemistry to yield a stable triazole linkage with azide; copper is needed as a catalyst. With PEG chain embedded in the molecule, the hydrophilicity is greatly improved. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
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