CO00016 Midstream Cassette 140x15mm, blue w/lens

N-(Propargyl-PEG4-carbonyl)-N-bis(PEG1-acid) is a crosslinker that can react with azide compounds or biomolecules via copper catalyzed Click Chemistry to form a stable triazole linkage. The terminal carboxylic acids can react with primary amino groups in the presence of activators (e.g. HATU) to form a stable amide bond.

N-(Propargyl-PEG4-carbonyl)-N-bis(PEG1-acid) is a crosslinker that can react with azide compounds or biomolecules via copper catalyzed Click Chemistry to form a stable triazole linkage. The terminal carboxylic acids can react with primary amino groups in the presence of activators (e.g. HATU) to form a stable amide bond.

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.