Diazo Biotin-PEG3-alkyne is useful for introducing a biotin moiety to azide-containing biomolecules using Cu(I)-catalyzed Click Chemistry. The hydrophilic spacer arm provides better solubility to the labeled molecules in aqueous media. Diazo allows efficient release of captured biotinylated molecules from streptavidin using sodium dithionite (Na2S2O4). Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
Diazo Biotin-PEG3-alkyne is useful for introducing a biotin moiety to azide-containing biomolecules using Cu(I)-catalyzed Click Chemistry. The hydrophilic spacer arm provides better solubility to the labeled molecules in aqueous media. Diazo allows efficient release of captured biotinylated molecules from streptavidin using sodium dithionite (Na2S2O4). Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
DBCO-Acid is a non-activated building block and adds minimal spacer to modified molecules. In the presence of activators such as EDC or HATU, this reagent can be used to derivatize amine-containing molecules through a stable amide bond. DBCO is commonly used for copper-free Click Chemistry reactions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
DBCO-Acid is a non-activated building block and adds minimal spacer to modified molecules. In the presence of activators such as EDC or HATU, this reagent can be used to derivatize amine-containing molecules through a stable amide bond. DBCO is commonly used for copper-free Click Chemistry reactions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
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 |
The CloneSizer 100 bp DNA Ladder is prepared to ensure quality and batch-to-batch consistency. Our CloneSizer contains fourteen discrete fragments ranging from 100 bp to 2000 bp in 100 bp increments, double intensity reference bands at 500 and 1000 bp and an additional 2686 bp (pUC19) reference band for easy clone identification.
Contents:
1mL of premixed DNA ladder (0.5µg/10µL) in loading buffer (10mM EDTA, 10% glycerol, 0.015% bromophenol blue, and 0.17% SDS).
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CloneSizer 100 bp DNA Ladder (Cat# 11600) – 100 loads
Ladder Properties:
| Fragment | Size (bp) | Mass (ng) |
| 1 | 2686 | 72 |
| 2 | 2000 | 53 |
| 3 | 1500 | 41 |
| 4 | 1200 | 42 |
| 5 | 1000 | 56 |
| 6 | 900 | 30 |
| 7 | 800 | 29 |
| 8 | 700 | 25 |
| 9 | 600 | 25 |
| 10 | 500 | 52 |
| 11 | 400 | 19 |
| 12 | 300 | 20 |
| 13 | 200 | 19 |
| 14 | 100 | 17 |
Recommended Use:
Mix thoroughly. For best results, load 10µL of DNA ladder per well. For precise mass determination with a densitometer, stain gel after electrophoresis using 0.5µg/mL ethidium bromide for 30-40 minutes. The table above shows the size and mass for each band based on 10µL ladder per well.
Storage:
Stable at room temperature. For longer term storage, -20°C is recommended.
This ladder was standardized using 10µL of DNA per lane on a 0.8 cm thick, 13 x 15 cm, 1.0% agarose gel run in TAE buffer.
