t-Boc-N-Amido-PEG3-propargyl

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

Intended Use

Used for the selective isolation and culture of Listeria monocytogenes.

Principle and Interpretation

Columbia Blood Agar Base provide carbon and nitrogen sources, vitamins and growth factors，maintains balanced osmotic pressure; Listeria hydrolyzes aesculin and reacts with iron ions to form black 6,7-dihydroxycoumarin; lithium chloride and other antibiotics can inhibit Gram-negative bacteria and most Gram-positive bacteria; agar is the coagulant of the culture medium.

Formulation

Ingredients	/liter
Columbia blood agar base	39.0 g
Aesculin	1.0 g
Ferric ammonium citrate	0.5 g
Lithium chloride	15.0 g
pH7.0±0.2 at 25°C

Preparation

Weigh 55.5 g of this product, add 1000 mL of distilled water or deionized water, stir, heat and boil until completely dissolved, divide into Erlenmeyer bottles, sterilize at 121℃ for 15 min. Cool to about 50℃, add 1 bottle of SR0500 supporting reagent A and 1 bottle of B per 100 mL, mix well and set aside.

Quality Control

The following quality control strains were inoculated and cultured at 35-37℃ for 24h. The results are as follows:

Quality control strains	Growth
Listeria monocytogenes ATCC19115	Gray-green colonies with a black depression in the center and black surrounding
Enterococcus faecalis ATCC29212	–
Escherichia coli ATCC25922	–

Storage and Shelf Life

2-30℃，Keep container tightly closed, avoid direct sunlight.

Use before expiry date on the label.

 Precautions

1. When weighing the dehydrated medium, please wear masks to avoid causing respiratory system discomfort

2. Keep container tightly closed after using to prevent clumping.

Waste Disposal

Microbiological contamination was disposed by autoclaving at 121°C for 30 minutes.

Intended Use Used for the selective isolation and culture of Listeria monocytogenes. Principle and Interpretation Columbia Blood Agar Base provide carbon and nitrogen sources, vitamins and……

N-Me-N-bis(PEG2-propargyl) has two alkyne groups which can participate in Click Chemistry reactions with azide-bearing compounds or biomolecules; copper is required for catalyzation. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

N-Me-N-bis(PEG2-propargyl) has two alkyne groups which can participate in Click Chemistry reactions with azide-bearing compounds or biomolecules; copper is required for catalyzation. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.