DBCO-amine TFA salt is a simple building block containing a DBCO moiety and an amine group. Amine is very reactive with NHS ester. DBCO is commonly used for copper-free Click Chemistry reactions.

DBCO-amine TFA salt is a simple building block containing a DBCO moiety and an amine group. Amine is very reactive with NHS ester. DBCO is commonly used for copper-free Click Chemistry reactions.

The series of DNA Size Selection Kits (Magnetic Beads) were developed for DNA size selection using magnetic beads. A total of 11 kits are available, with different selection ranges spanning from 50 bp to over 10 kb. The kits provide a simple and quick approach for the enrichment of a specific range of DNA fragments. The kit workflow allows double-sided or single-sided size selection for specific size cutoffs.

Gel images of different ranges of size selection. Sheared human genomic DNA was used as input.

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DNA size selection is a selective capture of DNA fragments of a specific range of size for next-generation sequencing (NGS) library preparations, PCR, ChIP assay, DNA ligations, endonuclease digestions, adapter removal, and other genomics and molecular biology applications. DNA size selection is preferred after NGS library prep in most of the cases. The NGS library preparation is related to the quality of the sequencing data. Precise NGS library size selection can increase sequencing efficiency, improve data quality, and reduce costs.

There are two types of sequencing technologies: short-read sequencing and long-read sequencing. Short-read sequencing uses DNA libraries that contain small insert DNA fragments of similar sizes, usually several hundred base pairs. The sequencing efficiency can be improved if the DNA size selection is in the right range. Cat.# 20104S and 20104L are the best kits for NGS library size selection of illumina paired-end 100 (PE100) sequencing with 100-200 bp library inserts; Cat.# 20105S and 20105L are the best kits for NGS library size selection of illumina paired-end 150 (PE150) sequencing with 150-300 bp library inserts; and Cat.# 20106S and 20106L are the best kits for NGS library size selection of illumina paired-end 300 (PE300) sequencing with 300-600 bp library inserts.

Long-read sequencing uses a large DNA fragment as input and makes very long reads. Usually, library size selection is preferred to remove smaller fragments. Cat.# 20110S and 20110L are the best kits for long-read sequencing size selection with DNA sizes >5 kb, and Cat.# 20111S and 20111L are the best kits for long-read sequencing size selection with DNA sizes >10 kb.

The magnetic beads technology uses paramagnetic particles, also known as SPRI (Solid Phase Reversible Immobilization) beads, to bind DNA reversibly and selectively. DNA fragments can be size-selected and purified by changing the properties of the magnetic beads or SPRI beads. The magnetic beads can easily separate the beads-binding DNA from the contaminants and unwanted components in the samples. The samples after DNA size selection are free of contaminants such as buffer components, enzymes, proteins, salts, dNTPs, primers, and adapters. Our proprietary magnetic beads reagents improve yield, selectivity, and reproducibility.

Specific DNA fragments at a certain length range can be purified simply using magnetic separation with different beads components, avoiding tedious and time-consuming gel extraction and column-based purification. The magnetic beads method is popular for common DNA size selection, including library size selection. The first beads-binding step, referred to as the right-side clean-up, removes large DNA fragments. The large DNA fragments are bound to the beads and are discarded. The desired DNA fragments in the supernatant are transferred to a new well, and new beads are added to the supernatant for the second beads-binding, referred to as the left-side clean-up. The double-size selected DNA fragments are eluted after ethanol rinsing.

DNA size selection with dual clean-ups.

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A single clean-up is needed for DNA size selection with large fragments. In this case, only the large DNA fragments are bound to the beads. The selected larger DNA fragments are eluted after ethanol rinsing.

DNA size selection with single clean-up for >5 kb and >10 kb DNA.

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Features of DNA size selection and library size selection

• • • High specificity and high recovery of size selection
    • 11 selection ranges are available, including 5 ranges for NGS library size selection
      • • 50-100 bp
        • 100-200 bp
        • 200-500 bp
        • 250-350 bp: ideal for illumina PE100 sequencing
        • 300-450 bp: ideal for illumina PE150 sequencing
        • 450-750 bp: ideal for illumina PE300 sequencing
        • 500-1000 bp
        • 1-3 kb
        • 1-5 kb
        • >5 kb: ideal for long-read sequencing
        • >10 kb: ideal for long-read sequencing
    • Fast and simple
      • • 20-min protocol
        • No gel purification required
        • No columns required
        • No centrifugation required
    • Efficient removal of contaminants and unwanted components

Introduction

Free-circulating nucleic acids, such as tumor-specific extracellular nucleic acid fragments and mRNAs in the blood or fetal nucleic acids in maternal blood, are present in serum or plasma usually as short fragments, <1000bp(Nucleic Acid). The HiPure Circulating Nucleic acid Micro Kit enables efficient purification of these circulating nucleic acids from human plasma, serum, or urine. Samples can be either fresh or frozen (provided that they have not been frozen and thawed more than once). Free-circulating cell-free DNA, RNA or viral nucleic acids are eluted in Nuclease Free Water, ready for use in amplification reactions or storage at -30 to -15°C. Purified nucleic acids are free of proteins, nucleases, and other impurities.

Details

Specifications

Features	Specifications
Main Functions	Isolation circulating DNA from 0.6ml 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	0.6ml
Elution volume	≥30μl
Time per run	≤40 minutes
Liquid carrying volume per column	800μl
Binding yield of column	100μg

Principle

This product is based onsilica column purification. The sample is lysed and digested with lysate andprotease, DNA is released into the lysate. Transfer to an adsorption plate andfilter column. Nucleic acid is adsorbed on the membrane, while protein is notadsorbed and is removed with filtration. After washing proteins and otherimpurities, Nucleic acid was finally eluted with low-salt buffer.

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 recoveredat the level of PG by silica gel column purification

Kit Contents

Contents	D318002	D318003
Purification Times	50 Preps	250 Preps
Buffer ACL	40 ml	200 ml
Buffer DCW1	22 ml	110 ml
Buffer DCW2*	20 ml	2 x 50 ml
Proteinase K	34 mg	180 mg
Protease Dissolve Buffer	1.8 ml	10 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	5 x 100

Storage and Stability

Proteinase K, Carrier RNAshould be stored at 2-8°C upon arrival. However, short-term storage (up to 12weeks) at room temperature (15-25°C) does not affect their performance. Theremaining kit components can be stored dry at room temperature (15-25°C) andare stable for at least 18 months under these conditions. The entire kit can bestored 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.

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