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 |
Okadaic Acid (OA) is a one of the diarrhetic shellfish poisons (DSP) produced by dinoflagellate genera Dinophysis and Prorocentrum. There are several chemically different toxins associated with DSP.
They are lipophilic and polyether compounds and can be divided into three main groups:
In humans, DSP causes dose-dependent symptoms of diarrhea, nausea, and vomiting. The action levels established by the FDA for OA is 200ppb. The EU has established a level of 160 ppb of OA or its equivalent.
The Attogene Okadaic acid ELISA kit enables international and government regulatory agencies, food manufacturers and processors, as well as quality assurance organizations to detect OA in food, feed, fish, and environmental samples of concern.
Okadaic acid is the causative agent of Diarrhetic Shellfish Poisoning (DSP).
FDA and EPA Safety Levels in Regulations and Guidance – 0.16 mg/kg for Clams, mussels, oysters, and whole and roe-on scallops, fresh, frozen, or canned. – National Shellfish Sanitation Program Guide for the Control of Molluscan Shellfish.
Competitive ELISA for the quantitative analysis of Okadaic Acid (DSP)
Format: 96-well microtiter plate (12 test strips of 8 wells)
Okadaic acid is a potent neurotoxin and phosphatase inhibitor from dinoflagellate black sponges that are associated with seafood poisonings.
Propargyl-PEG10-NHS ester serves as a bifunctional PEG linker with a terminal propargyl that participates in click reactions with azide-bearing moieties and an NHS ester group that reacts readily and efficiently with amine-bearing molecules. The PEG10 units improve the water solubility of the molecule. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
Propargyl-PEG10-NHS ester serves as a bifunctional PEG linker with a terminal propargyl that participates in click reactions with azide-bearing moieties and an NHS ester group that reacts readily and efficiently with amine-bearing molecules. The PEG10 units improve the water solubility of the molecule. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.