Overview

• Protocol optimized for DNA isolated from a diversity of samples including stool, soil, water, saliva, plant, urine, skin, and more
• Simple and quick workflow: library could be prepared in less than 5 hours
• Component of Norgen’s metagenomics workflow
• A single NGS run can be prepared with up to 384 unique dual-index libraries

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.

Details

Supporting Data

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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.

Magnetic Beads (microRNA & Oligo Purification)

Solid Phase Reversible Immobilization magnetic beads are often used for DNA purification because they are simple, fast, and effective. The beads are paramagnetic particles coated with carboxyl groups that reversibly bind to nucleic acid. However, Standard SPRI beads can only purify DNA/RNA fragments that are 100 base pairs or longer. DNA/RNA fragments shorter than 100 base pairs are not effectively recovered by SPRI beads. We have developed Magnetic Beads (microRNA & Oligo Purification) to solve the problem.

With our proprietary beads technology, the beads overcomes the hurdle of the short DNA/RNA recovery problem. The magnetic beads is ideal for microRNA purification, oligo purification, short DNA/RNA purification, and removing impurities and unwanted components such as dNTPs, detergents, salts, proteins, and other contaminants effectively. The magnetic bead reagents are RNase-free and can be used for both DNA and RNA applications.

Our magnetic beads are optimized for microRNA purification, oligo purification, and DNA/RNA purification. The fragments can be as short as 20 bases, such as microRNA, tRNA, dsDNA fragments 20 bp or longer, ssDNA fragments 20 nt or longer, RNA fragments 20 nt or longer, DNA/RNA hybrid fragments 20 bp or longer, and oligos and chimeric oligos 20 nt or longer. Purified short DNA and RNA fragments are ideal for applications requiring high-quality fragments, as the fragments are free of impurities and contaminants.

Comparison of short DNA fragments recovery. BioDynami 20 bp DNA ladder (Cat. # 10002) was used as DNA input. Ampure XP beads, BioDynami beads (DNA & RNA purification) and BioDynami beads (microRNA & Oligo purification) were tested. Only BioDynami beads (microRNA & Oligo purification) successfully recovered DNA fragments between 20-100 bp.

Recovery of DNA and RNA oligos with BioDynami magnetic Beads (microRNA & Oligo Purification). 20 nt of DNA oligos and RNA oligos were used as input. Input and recovered oligos were quantified with BioDynami ssDNA Quantification kit (Cat. # 40043) and BioDynami RNA Quantification kit (Cat. # 40044).

Features

• Effective purification of short DNA and RNA samples
  • • microRNA
    • tRNA
    • dsDNA fragments 20 bp or longer
    • ssDNA fragments 20 nt or longer
    • RNA fragments 20 nt or longer
    • DNA/RNA hybrid fragments 20 bp or longer
    • Oligo and chimeric oligo 20 nt or longer
• Removal of impurities and unwanted reaction components
• Compatibility: Works with both manual and automated procedures

Bacterial DNA Extraction Kit (Magnetic Beads)

The Bacterial DNA Extraction Kit (Magnetic Beads) was developed for the bacterial genomic DNA extraction from bacterial cultures directly using magnetic beads from a wide variety of gram-negative and gram-positive bacterial species, as well as yeast and other fungi. With our proprietary magnetic beads technology, the kit eliminates the tedious centrifuge steps for columns. The kit provides a reliable and simple approach for high-quality bacterial DNA isolation with fast magnetic response time and high binding capacity.

Bacteria are the most diverse and abundant small single-celled organisms and are vital to the planet’s ecosystems. Some bacterial species are pathogens that can cause a variety of diseases to humans and animals. Besides their ecological and biomedical importance, bacteria are also used in biotech and pharmaceutical applications such as production of enzymes, DNA preparation, biofuels, food research, and chemical production.

Bacterial cells are grown to log-phase and the culture is lysed directly with a lysis buffer, then mixed with beads to bind genomic DNA. After wash steps, genomic DNA is eluted in Low TE or TE Buffer. The isolated genomic DNA with the magnetic beads is free of contamination such as RNA, proteins, salts, and other impurities. Bacterial DNA extracted using the kit is suitable for downstream applications such as qPCR, PCR, DNA sequencing, Southern Blotting, molecular cloning, DNA hybridization, restriction enzymatic digestion, and Next-generation Sequencing (NGS) etc.

Features

• 100% centrifuge-free
  • Bacterial cultures can be used directly without centrifuge to pellet the bacteria
• Simple magnetic beads method
  • No centrifuge needed
  • No column needed
  • No vacuum needed

Purified genomic DNA from various bacteria were isolated with the Bacterial DNA Extraction Kit. A portion of the extracted genomic DNA was loaded on a 1% agarose gel. DNA ladder: BioDynami 1 kb Plus DNA Ladder (Cat.# 10005L).

Amine-Tri(3-methoxypropanamide-PEG10-Propargyl) Methane HCl salt is reactive with azide-bearing compounds or biomolecules via copper catalyzed azide-alkyne Click Chemistry to yield a stable triazole linkage. The amino group (NH2) is reactive with carboxylic acids, activated NHS esters, carbonyls (ketone, aldehyde), etc. Reagent grade, for research use only.

Amine-Tri(3-methoxypropanamide-PEG10-Propargyl) Methane HCl salt is reactive with azide-bearing compounds or biomolecules via copper catalyzed azide-alkyne Click Chemistry to yield a stable triazole linkage. The amino group (NH2) is reactive with carboxylic acids, activated NHS esters, carbonyls (ketone, aldehyde), etc. Reagent grade, for research use only.

TD5A Low Speed Centrifuge

Features:

1. Microprocessor control, digital display, touch panel, parameters in running can be edited and change to display other running parameter and RCF.

2. Brushless frequency motor with simpler construction, more reliable performance, longer life and quietly running.

3. Automatic lid lock, super speed, over temperature protection and imbalance protection. The centrifuge body is made of high quality steel, safe and reliable.

4. Adapters are available by experiment requirements.                          

5. 3 tiers protection steel cover, safe and reliable.

6. TUV CE EMC certificate.

TD5A Technical Parameter：

Matched Rotor for TD5A

TD5A centrifuge Max Capacity 4x300ml swing out rotor, max speed 5000rpm, can hold 96 vacuum tubes