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

Description

Nucleic acid testing (NAT) is the method of choice for detection and quantification of a wide range of micro organisms. Primerdesign manufactures and supplies high quality quantitative real-time PCR kits for the detection and simultaneous quantification of numerous significant pathogens . A copy number standard curve is provided for quantification and an the internal extraction template (DNA or RNA), controls for the quality of the nucleic acid extraction and eliminates false negative results.

The kit is designed with the broadest possible detection profile to ensure that all clinically relevant strains and subtypes are detected. Target sequences are selected by working with data from key opinion leaders in the field. Multiple sequence alignments and unprecedented real-time PCR expertise in design and validation ensure the best possible kit.

Details of the target and priming specificity are included in the individual handbooks above.

Packaged, optimised and ready to use. Expect Better Data.

Exceptional value for money
Rapid detection of all clinically relevant subtypes
Positive copy number standard curve for quantification
Highly specific detection profile
High priming efficiency
Broad dynamic detection range (>6 logs)
Sensitive to < 100 copies of target Accurate controls to confirm findings 50 & 150 reactions

Introduction

MagPure Stool DNA Kit is specially designed for high throughput DNA extraction from stool samples. It can get high purity microbial DNA from stool samples (≤200mg). This kit is based on magnetic beads purification and unique inhibiting factors adsorption technology, no phenol-chloroform extraction or alcohol precipitation. It can adsorb humic acid and other inhibiting factors in the solution efficiently. DNA can be directly used for downstream applications such as PCR, Viral DNA testing, bacterial DNA testing, etc.

Details

Specifications

Features	Specifications
Main Functions	Isolation total DNA from 100-150mg stool samples
Applications	PCR, southern blot and enzyme digestion, etc.
Purification technology	Magnetic beads technology
Process method	Manual or automatic
Sample type	Stool
Sample amount	100-150 mg
Elution volume
Time per run

Principle

This product is based on the purification method of high binding magnetic particles. The sample is lysed and digested under the action of lysate and Lysozyme. DNA is released into the lysate. After adding magnetic particles and binding solution, DNA will be adsorbed on the surface of magnetic particles, and impurities such as proteins will be removed without adsorption. The adsorbed particles were washed with washing solution to remove proteins and impurities, washed with ethanol to remove salts, and finally DNA was eluted by Elution Buffer.

Advantages

• Fast – several samples can be extracted in 60 minutes (after digestion)
• High purity – unique adsorbent can completely remove inhibitory factors
• High recovery – DNA can be recovered at the level of PG

Kit Contents

Contents	D636401	D636402	D636404
Purification Times	48 Preps	96 Preps	400 Preps
MagPure Particles N	1.7 ml	3.4 ml	14 ml
2ml Bead Tubes	48	96	400
RNase A	10 mg	20 mg	75 mg
Proteinase K	24 mg	48 mg	180 mg
Protease Dissolve Buffer	3 ml	5 ml	20 ml
Buffer ATL	60 ml	110 ml	300 ml
PVP-10	1.2 g	2.2 g	6 g
Buffer PCI	50 ml	100 ml	300 ml
Buffer MLE	30 ml	60 ml	180 ml
Buffer GW1*	22 ml	44 ml	132 ml
Elution Buffer	20 ml	20 ml	60 ml

Storage and Stability

MagPure Particles, RNase A and Proteinase K should be stored at 2-8°C upon arrival. However, short-term storage (up to 24 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.

MagPure Stool DNA Kit is specially designed for high throughput DNA extraction from stool samples. It can get high purity microbial DNA from stool samples (≤200mg). This kit is based on magnetic beads purification and unique inhibiting factors adsorption technology, no phenol-chloroform extraction or alcohol precipitation. It can adsorb humic acid and other inhibiting factors in the solution efficiently. DNA can be directly used for downstream applications such as PCR, Viral DNA testing, bacterial DNA testing, etc.