DOTA-PEG5-C6-DBCO is a PEG linker containing DOTA and DBCO moieties. DBCO is used in Click Chemistry reactions due to its high strain energy. The DOTA group can be ionized and is susceptible for chelating di- and trivalent cations. DOTA can also be used for imaging diagnostic techniques. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

DOTA-PEG5-C6-DBCO is a PEG linker containing DOTA and DBCO moieties. DBCO is used in Click Chemistry reactions due to its high strain energy. The DOTA group can be ionized and is susceptible for chelating di- and trivalent cations. DOTA can also be used for imaging diagnostic techniques. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

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.

Introduction

Free-circulating nucleic acids, such as tumor-specific extracellular DNA fragments and mRNAs in the blood or fetal nucleic acids in maternal blood, are present in serum or plasma usually as short fragments, <1000bp (DNA). HiPure Circulating DNA Mini Kit enables efficient purification of these circulating nucleic acids from human plasma, serum, or urine. Samples can be fresh or frozen (provided that they have not been frozen and thawed more than once).

Details

Specifications

Features	Specifications
Main Functions	Isolation circulating DNA from 1ml plasma, serum, body fluids
Applications	qPCR, liquid or solid chip analysis, hybridization and SNP detection, etc.
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	Serum, plasma and other cell-free fluid samples
Sample amount	1ml
Elution volume	≥30μl
Time per run	≤50 minutes
Liquid carrying volume per column	800µl
Binding yield of column	100µg

Principle

This product is based on silica gel purification. The sample is lysed and digested with lysate and protease, DNA is released into the lysate. Transfer to an adsorption plate and filter column. Nucleic acid 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 (10 Mm Tris,pH 8.0).

Advantages

• High yield – most optimal process, free DNA (>50bp) can be obtained to the maximum extent
• High concentration – low elution volume, ensuring high nucleic acid concentration
• High purity – low alcohol binding method, completely removing inhibitor and protein pollution
• High recovery – DNA can be recovered at the level of PG by silica gel column purification

Kit Contents

Contents	D318102	D318103
Purification Times	50 Preps	250 Preps
Buffer ACL	50 ml	250 ml
Buffer ACB*	60 ml	300 ml
Buffer DCW1*	22 ml	88 ml
Buffer DCW2*	10 ml	50 ml
Proteinase K	120 mg	540 mg
Protease Dissolve Buffer	10 ml	30 ml
Carrier RNA	110 μg	310 μg
Nuclease Free Water	10 ml	30 ml
HiPure CFDNA Mini Columns	50	250
2 ml Collection Tubes	100	500

Storage and Stability

Proteinase K and carrier RNA 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 remainingkit 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 shouldbe redissolved before use. Make sure that all buffers are at room temperature when used.

Free-circulating nucleic acids, such as tumor-specific extracellular DNA fragments and mRNAs in the blood or fetal nucleic acids in maternal blood, are present in serum or plasma usually as short fragments,