The NGS DNA Fragmentation & Library Prep Kit (illumina and MGI Platforms) was developed for construction of high quality libraries for next generation sequencing. The kit uses intact genomic DNA (EDTA-free DNA or DNA resuspended in TE buffer) as input DNA without an additional DNA fragmentation step. Our technology provides a fast and simple workflow. DNA libraries can be generated around 2 hours with only 10 min hands-on time. Library multiplexing is possible.

NGS DNA Fragmentation & Library Prep Kit Workflow

The incorporation of DNA fragmentation in the kit makes it possible to directly use intact genomic DNA as input DNA without the need of mechanical DNA shearing or enzymatic DNA fragmentation. The NGS DNA Fragmentation & Library Prep Kit does not generate sequencing bias as compared to library using mechanical sheared DNA as input. Sequence coverage is also consistent between enzymatic shearing and mechanical shearing. The library size is inversely correlated with the incubation time of step 1 at 20°C.

Three index types are available for the kit of the illumina platform:

Non-index (Cat.# 30026): Libraries do not have index.

Index (Cat.# 30028): Each of the index primers contains a unique index sequence of 6 bases. Library multiplexing for 48 samples is possible. Index information can be downloaded here.

Unique dual index (Cat.# 30030): Library multiplexing for 96 samples is possible. With the unique features of our 4-Base Difference Index System, the index sequence is 8-base long and each index has 4 bases different from others. Our unique dual index primers effectively identify sequencing errors such as index hopping, mis-assignment of reads, and de-multiplexing errors etc. The unique dual index primers set consists of 96 pre-mixed unique pairs of  index primers. Index information can be downloaded here.

Indexes are available for the MGI platform kits (Cat.# 34028).

Kit features:

• • • 1.5-hour protocol from intact genomic DNA to NGS library
    • Intact genomic DNA as input, DNA fragmentation is not needed.
    • Works with both EDTA-free DNA and DNA resuspended in TE buffer
    • Simple workflow: Less steps
    • Guaranteed quality: Higher library conversion efficiency

Library conversion efficiency: 100 ng, 300 ng and 500 ng of intact genomic DNA were used as input.

The library size is inversely correlated with the incubation time of step 1 at 20°C.

NGS data comparison: enzymatic shearing versus mechanical shearing

Enzymatic shearing
• DNA shearing and library prep: BioDynami NGS DNA Fragmentation & Library Prep Kit
Mechanical shearing
• DNA shearing: Covaris sonication
• Library prep: BioDynami NGS DNA Library Prep Kit.

The NGS DNA Fragmentation & Library Prep Kit (illumina and MGI Platforms) was developed for construction of high quality libraries for next generation sequencing. The kit uses intact genomic DNA (EDTA-free DNA or DNA resuspended in TE buffer) as input DNA without an additional DNA fragmentation step. Our technology provides a fast and simple workflow. DNA libraries can be generated around 2 hours with only 10 min hands-on time. Library multiplexing is possible.

Product advantages

1. The instantaneous centrifugal function rises to the maximum speed for about 6 seconds, allowing for rapid detachment of hanging droplets from the wall;

2. Faster acceleration and deceleration rate, required to complete the experiment in a shorter time;

3. Door cover protection, overspeed and imbalance detection system, capable of real-time monitoring of  the centrifugal process, ensuring the safe operation    of the  instrument; When the operation ends, an error occurs, or an imbalance occurs, the sound signal prompts, and the operation stops at the same time. The LCD displays the result code

Host Parameters

The micro porous plate centrifuge adopts international advanced design concepts and manufacturing technology, with a smooth and beautiful appearance, compact and stable structure, adhering to the advantages of high universality and easy operation. It also has the functional characteristics of “smooth start” and “smooth braking”; This micro porous plate centrifuge is very suitable for 96 or 384 well and small capacity micro porous plates, as well as various standard PCR micro porous plates with or without skirts .

Introduction

This product provides an easy-to-use workflow for selective isolation of bacterial DNA from samples that are intrinsically rich in host DNA, such as body fluids or swabs. The method is specific for the identification of intact bacteria so it prevents false results due to nucleic acids from dead bacteria. The Kit allows isolation of enriched bacterial DNA suitable for a variety of applications, including qPCR and whole metagenome or 16S rRNA gene sequencing.

Details

Specifications

Principle

This product is based on silica Column purification. This product efficiently depletes human and animal host DNA and yields enriched bacterial DNA. An optimized combination of mechanical and chemical lysis allows efficient disruption of bacterial cells. Target DNA is adsorbed on the membrane, while protein is not adsorbed and is removed with filtration. After washing proteins and other impurities, nucleic acid was finally eluted with low-salt buffer (10mm Tris, pH9.0, 0.5mm EDTA).

Advantages

• High concentration – the host DNA is digested by dnase, without contamination of host DNA
• High recovery – DNA can be recovered at the level of PG
• Good repeatability – silica technology can obtain ideal results every time
• Wide applicability – it can be used in blood, tissue, intestinal contents and other samples

Kit Contents

Storage and Stability

DNase I and Proteinase K should be stored at 2–8°C upon arrival. However, short-term storage (up to 12 weeks) at room temperature (15–25°C) does not affect their performance. The remaining kit components can be stored dry at room temperature (15–25°C) and are stable for at least 18 months under these conditions. The entire kit can be stored at 2–8°C, but in this case buffers should be redissolved before use. Make sure that all buffers are at room temperature when used.

This product provides an easy-to-use workflow for selective isolation of bacterial DNA from samples that are intrinsically rich in host DNA, such as body fluids or swabs. The method is specific for the identification of intact bacteria so it prevents false results due to nucleic acids from dead bacteria. The Kit allows isolation of enriched bacterial DNA suitable for a variety of applications, including qPCR and whole metagenome or 16S rRNA gene sequencing.