Sample type purification kit guide
The 16S V3-V5 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-V5 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.
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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 V3-V5Library 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. 70500 (24 preps) | Cat. 70510, 70520, 70530, 70540 (96 preps) |
|---|---|---|---|
| Amplicon PCR (PCR 1) | MGX Master Mix | 330 µL | 1,320 µL |
| 16S V3-V5 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 |
Propargyl-PEG6-acid comprises propargyl and carboxylic acid functional groups. The acid group reacts with primary amines in the presence of activators (e.g. EDC, or HATU).The propargyl group reacts with azide-bearing compounds or biomolecules via copper catalyzed azide-alkyne Click Chemistry to yield a stable triazole linkage. The molecule has good solubility in aqueous environment. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
Propargyl-PEG6-acid comprises propargyl and carboxylic acid functional groups. The acid group reacts with primary amines in the presence of activators (e.g. EDC, or HATU).The propargyl group reacts with azide-bearing compounds or biomolecules via copper catalyzed azide-alkyne Click Chemistry to yield a stable triazole linkage. The molecule has good solubility in aqueous environment. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
Efficient, leaf punch tools for quick, precise field sampling. For use with the 3CR Bio Leaf Sample Collection Kit.
BASIC LEAF CUTTING TOOL
The 3CR Bio Basic Leaf Cutting Tool is a lightweight, easy-to-use, low-cost punch designed for precise sampling of thin or soft materials, ideal for fieldwork. This tool is expertly designed to core, punch, and retrieve samples from thin or soft source materials such as leaves. The tool comprises a convenient, hollow plastic pen-style holder, mounted with a durable stainless steel cutting tip at one end. It features an internal push-button-operated plunger for safe and efficient sample disc ejection. The tool produces a precise ¼ inch diameter disc, making it ideal for detailed sampling tasks. For best results, clean the cutting tip between each sample extraction by coring blank filter paper or rinsing with water or ethanol. This low-cost leaf punch is lightweight, easy to use, and perfect for fieldwork.
ADVANCED LEAF CUTTING TOOL
The 3CR Bio Advanced Leaf Cutting Tool is designed to streamline and enhance your sampling process. This high-precision leaf tissue punch produces a single ¼ inch diameter disc effortlessly. It features a unique mechanism that securely holds a standard snap-top or screw-top 2 mL storage tube in a fixed position, allowing the sample to be received without any need for touching or manipulation. Ideal for fieldwork, this advanced leaf punch significantly reduces effort and time when performing multiple samplings, ensuring accuracy and efficiency in every use.
Efficient, leaf punch tools for quick, precise field sampling. For use with the 3CR Bio Leaf Sample Collection Kit.