DBCO-PEG4-PABA

Description

The EzRNA™ T7 High Yield RNA Synthesis Kit (Ψ-UTP) is a user-friendly product for enzymatic RNA production. The enzyme mix contains adequate amount of T7 RNA polymerase, pyrophosphatase, and RNase inhibitors for in vitro transcription (IVT). Along with 10X Transcription Buffer and NTP (Ψ) Premix, users can swiftly assemble IVT reactions without compromising RNA yield. The EzRNA™ T7 High Yield RNA Synthesis Kit (Ψ-UTP) allows for the attainment of approximately up to 150 µg RNA yield within 2 hours at 37°C.

Features

• High yield
• Versatile- suitable for short and long transcripts
• NTP premixed- Minimal pipetting and setup time
• Compatible with CleanCap® Reagent AG
• Lithium chloride included for RNA purification

Application

• Generation of RNA from T7 promoter-driven DNA sequences
• Suitable for subsequent cap-0 and cap-1 modification

Storage

-20°C for 12 months

The EzRNA™ T7 High Yield RNA Synthesis Kit (Ψ-UTP) is a user-friendly product for enzymatic RNA production. The enzyme mix contains adequate amount of T7 RNA polymerase, pyrophosphatase, and RNase inhibitors for in vitro transcription (IVT). Along with 10X Transcription Buffer and NTP (Ψ) Premix, users can swiftly assemble IVT reactions without compromising RNA yield. The EzRNA™ T7 High Yield RNA Synthesis Kit (Ψ-UTP) allows for the attainment of approximately up to 150 µg RNA yield within 2 hours at 37°C.

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

This product is suitable for rapid extraction of RNA from low RNA yield somples such as tissue (＜10mg), cells, bone marrow, fresh blood, and other clinical samples. RNA can be used directly for RT-PCR, Real time PCR, NGS, Viral RNA detection and so on.

Details

Specifications

Features	Specifications
Main Functions	Isolation total RNA from tissue, cell, whole blood
Applications	RT-PCR, cDNA synthesis, second generation sequencing
Purification method	Polydisperse magnetic beads
Purification technology	Magnetic beads technology
Process method	Manual or automatic
Adaptive instrument	Nucleic acid extractor, pipetting workstation
Sample type	Tissues, cells, lymphocytes and other clinical sample
Sample amount	Cells grown in suspension: ≤3 x 106Animal tissue: ≤10mgPlant tissue: ≤30mgWhole blood: 0.5~1.0 ml fresh blood or bone marrow and fresh blood mixture

Kit Contents

Contents	IVD3020B-96	IVD3020B
Purification Times	96 Preps	200 Preps
MagPure Particles N	2.5 ml	5 ml
DNase I	2 x 600 µl	4 x 600 µl
DNase Buffer C	60 ml	120 ml
Buffer RLC	60 ml	120 ml
Buffer MCB*	60 ml	150 ml
Buffer GW1*	44 ml	110 ml
Buffer MW2*	50 ml	100 ml
RNase Free Water	20 ml	60 ml

Storage and Stability

DNase I should be shipped with ice pack and stored at -20°C after arrival. MagPure Particles N should be stored at 2–8°C for long time storage. The remaining kit components can be stored at room temperature(15–25°C) for up to18 months under these conditions.

This product is suitable for rapid extraction of RNA from low RNA yield somples such as tissue (＜10mg), cells, bone marrow, fresh blood, and other clinical samples. RNA can be used directly for RT-PCR, Real time PCR, NGS, Viral RNA detection and so on.