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 |
For rapid, sensitive and accurate screening of potential Tyrosinase inhibitors
Tyrosinase (EC 1.14.18.1) is a copper-binding enzyme that is expressed across a vast range of species ranging from bacteria and fungi to mammals. It is involved in two sequential reactions of the melanin synthesis pathway: first being the hydroxylation of a monophenol and second the conversion of an ortho-diphenol to a quinone. Quinone then undergoes a series of reactions including polymerization to form melanin.
Tyrosinase is of great interest to the agriculture industry since it causes browning of fruits, vegetable, and mushrooms, as well as to the cosmetic industry as it causes skin darkening. Development and screening of tyrosinase inhibitors, therefore, is very useful for conditions such as hyperpigmentation and melasma. Tyrosinase activity is significantly increased in melanoma. Therefore, the detection of tyrosinase activity could be promising as a specific diagnostic test for melanoma and may be useful in monitoring patient response to melanoma treatments.
This Tyrosinase Activity Assay Kit is a simple one-step, plate-based assay for the measurement of tyrosinase activity in biological samples. In this assay, tyrosinase catalyzes the conversion of a phenolic substrate to a Quinone intermediate, which reacts with the tyrosine enhancer forming a highly stable chromophore with absorbance at 520 nm. The assay can detect as low as 30 μU Tyrosinase in biological samples.
Highly reproducibility
Highly Sensitive Assay to Screen for Tyrosinase Activity
Stable formulation of ready to use Reaction Facilitator (tyrosinase)
Opentrons Flex Magnetic Block (GEN 1) is a 96-well magnetic well plate holder containing static ring magnets, specifically designed for use with the Flex Robot. Use it with the Flex Gripper and skirted 96-well PCR or deep-well plates to facilitate quick and efficient separation using magnetic beads. This module is not compatible with the OT-2.
Opentrons Flex Magnetic Block (GEN 1) is a 96-well magnetic well plate holder containing static ring magnets, specifically designed for use with the Flex Robot. Use it with the Flex Gripper and skirted 96-well PCR or deep-well plates to facilitate quick and efficient separation using magnetic beads. This module is not compatible with the OT-2.