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

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Library size selection is an enrichment of a specific range of library sizes for NGS library preparations. 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, or SPRI (Solid Phase Reversible Immobilization) beads, is well used for the purification of DNA due to their reversible DNA binding. The NGS library can be size-selected by the magnetic beads or SPRI beads. The properties of the magnetic beads can be changed for a specific range of DNA binding. The contaminants and other unwanted components in the libraries can also be removed during size selection.

Specific ranges of NGS libraries can be selected using magnetic beads with different buffer compositions. The first DNA-beads binding step, also called the right-side clean-up, removes large NGS library fragments. The large NGS library fragments that bind to the beads are discarded with the beads pellet. The desired NGS library fragments in the supernatant are transferred to a new well, and new beads are added to the supernatant for the second beads-DNA binding, also called the left-side clean-up. After the rinsing step, the NGS library fragments with the dual selection are eluted in water or an appropriate buffer. The magnetic beads method has great advantages over time-consuming column purification and tedious gel-based purification.

NGS library size selection with dual clean-ups.

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Library size selection for long-read sequencing only requires a single clean-up. In this case, only the large library fragments are bound to the beads, while other small library fragments are discarded with the supernatant. The selected larger library fragments are eluted in water or an appropriate buffer after the rinsing step.

NGS library size selection with single clean-up for >5 kb and >10 kb libraries.

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Features of NGS library size selection

• • • 5 ranges for NGS library size selection
      • • illumina PE100 sequencing: 250-350 bp
        • illumina PE150 sequencing: 300-450 bp
        • illumina PE100 sequencing: 450-750 bp
        • Long-read sequencing: >5 kb
        • Long-read sequencing: >10 kb
    • Rapid protocol: only 20 minutes
    • Simple workflow
      • • No columns required
        • No gel purification required
        • No centrifugation required

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,

Description

• Description

Aflatoxin is the most common food toxin that is harmful to human and animal health. The most frequent aflatoxins are B1, B2, G1, and G2, which can affect the body through respiratory, mucosal, or cutaneous routes, causing an excessive inflammatory response. Aflatoxin can infect crops during their growing stages or even after they are harvested. It mainly targets the liver and can impair the effectiveness of immunization in children, increasing the risk of infection. Aflatoxin detection and quantification in food and feed is a critical part of food and feed safety concerns.

Aflatoxin is the most common food toxin that is harmful to human and animal health. The most frequent aflatoxins are B1, B2, G1, and G2, which can affect the body through respiratory, mucosal, or cutaneous routes, causing an excessive inflammatory response. Aflatoxin can infect crops during their growing stages or even after they are harvested. It mainly targets the liver and can impair the effectiveness of immunization in children, increasing the risk of infection. Aflatoxin detection and quantification in food and feed is a critical part of food and feed safety concerns.