Bis-propargyl-PEG18 has two propargyl groups at both ends. These propargyl groups reacts with azide compounds or biomolecules to form stable triazole linkages under the catalyzation of copper. The hydrophilic PEG units enhance the solubility of the molecule in aqueous environment. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
Bis-propargyl-PEG18 has two propargyl groups at both ends. These propargyl groups reacts with azide compounds or biomolecules to form stable triazole linkages under the catalyzation of copper. The hydrophilic PEG units enhance the solubility of the molecule in aqueous environment. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
Sample type purification kit guide
The 16S V3-V4 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-V4 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.
| 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-V4 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. 70400 (24 preps) | Cat. 70410, 70420, 70430, 70440 (96 preps) |
|---|---|---|---|
| Amplicon PCR (PCR 1) | MGX Master Mix | 330 µL | 1,320 µL |
| 16S V3-V4 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 |
This kit provides a convenient and rapid method to purify total RNA from small amounts of stool samples. All types of stool samples can be processed with this kit, including animal fecal samples, manure and samples collected using Norgen’s Stool Nucleic Acid Collection and Transport Devices Dx (Cat. Dx45660). The kit removes all traces of humic acids using rapid and simple spin column procedures. Bead tubes are also provided for effective homogenization of stool. The kit purifies all sizes of RNA, from large mRNA and ribosomal RNA down to microRNA and small interfering RNA. Both host and microbial RNA is recovered. The protocol does not rely on the use of phenol or chloroform, thereby providing a user friendly procedure and allowing high-throughput analysis. The purified RNA is of the highest integrity, and can be used in a number of downstream applications including real time PCR and reverse transcription PCR for gene expression analysis. The procedure can be completed in approximately 30 minutes.
NOTE: This product is not available for sale in the United States.
Figure 1 / 3
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| Kit Specifications | |
| Maximum Stool Input | 200 mg |
| Type of Stool Processed | Frozen and fresh stool |
| Maximum Column Binding Capacity | 50 μg |
| Maximum Column Loading Volume | 650 μL |
| Time to Complete 10 Purifications | 30 minutes |
Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment.
| Component | Cat. Dx49500 (50 preps) |
|---|---|
| Lysis Solution | 60 mL |
| Wash Solution | 22 mL |
| Elution Buffer | 6 mL |
| Bead Tubes | 50 |
| Spin Columns | 50 |
| Collection Tubes | 50 |
| Elution Tubes (1.7 mL) | 50 |
| Product Insert | 1 |
Endonucleases DNA-specific, dsDNase
Double-Strand Specific dsDNase (dsDNase) is ideal for fast and effective removal of contaminating DNA from PCR master mixes.
Taq polymerases are commonly contaminated by bacterial DNA. This is a problem in PCR based bacterial typing and detection as it might cause false positive results. The unique properties of dsDNase make it suited for removal of contaminating DNA from PCR master mixes prior to addition of DNA template.
In figure 1, a PCR master mix was treated with different amounts of dsDNase before performing a qPCR to measure the contaminating bacterial DNA in the master mix. ArcticZymes dsDNase effectively removed contaminating DNA below known levels of the assay detection limits.
The dsDNase from Arctic shrimp (Pandalus borealis) is recombinantly produced in Pichia pastoris. It cleaves phosphodiester linkages in DNA to yield oligonucleotides with 5’-phosphate and 3’-hydroxyl termini.
The specific activity is estimated to be 30 times higher than that of bovine DNase I. In the presence of magnesium as only divalent cation and using oligos as a substrate, the activity towards dsDNA is 5000-fold higher than towards ssDNA.
The unique double strand-specificity allows specific degradation of dsDNA while leaving shorter ssDNA as primers and probes essentially intact. Easy inactivation by moderate heat (65°C) allows addition of DNA intended for analysis directly after removal of contaminating DNA.
Figure 1. The dsDNase effectively removes contaminated DNA
The dsDNase effectively removes contaminated DNA:
A PCR master mix was preincubated with various concentrations of dsDNase. After treatment, no DNA was amplified in non-template controls.
Nucleic acid specificity has been tested towards double- and single-stranded DNA and RNA oligonucleotides. The specificity of dsDNase towards the substrate has been measured using 15-mer oligonucleotides with FAM at 5′ and DarkQuencher® 3′ (Eurogentec). The fluorescence is proportional to enzyme activity. Assay conditions: 25 mM Tris pH 7.5, 5 mM MgCl2, and 2 μM oligonucleotide.
Substrate Relative Activity
dsDNA 100%
ssDNA <0.03%
dsRNA <0.01%
ssRNA <0.01%
Double-Strand Specific dsDNase (dsDNase) is ideal for fast and effective removal of contaminating DNA from PCR master mixes.
Taq polymerases are commonly contaminated by bacterial DNA. This is a problem in PCR based bacterial typing and detection as it might cause false positive results. The unique properties of dsDNase make it suited for removal of contaminating DNA from PCR master mixes prior to addition of DNA template.
