NEST 2.0 mL Microcentrifuge Tube (500 count)

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

t-Boc-N-Amido-PEG4-propargyl is a propargyl linker with one side protected by Boc group. The propargyl group can form triazole linkage with azide-bearing compounds or biomolecules via copper catalyzed Click Chemistry. The Boc protected amine can be deprotected under mild acidic conditions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

t-Boc-N-Amido-PEG4-propargyl is a propargyl linker with one side protected by Boc group. The propargyl group can form triazole linkage with azide-bearing compounds or biomolecules via copper catalyzed Click Chemistry. The Boc protected amine can be deprotected under mild acidic conditions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

Description 

ExcelRT™ Reverse Transcription Kit is a complete, efficient and convenient kit to synthesize high quality first strand cDNA. This kit contains ExcelRT™ Reverse Transcriptase, which is able to synthesize the first strand cDNA at 37~50°C. The ExcelRT™ Reverse Transcriptase is a recombinant Moloney Murine Leukemia Virus (M-MLV) reverse transcriptase, which is designed to reduce RNase H activity and create better thermal stability. This kit also contains RNAok™ RNase Inhibitor, which is active against RNase A, RNase B, and RNase C. This product is supplied with oligo (dT)₂₀ and random hexamers, which are used to synthesize cDNA from poly(A) tailed mRNA and total RNA, respectively. 

Features

• Contains all components for reverse transcription 
• High yield 
• Thermostable, up to 50°C, during first strand synthesis 
• High processivity, generating cDNA up to 8 kb 
• Reduced RNase H ribonuclease activity 

Application

• Generation of first strand cDNA from total RNA or mRNA.
• Suitable for generating cDNA from RNA with strong secondary structure which can be reduced at temperature up to 50°C.

Storage

-20°C for 24 months 

High yield

High sensitivity

Thermostable, up to 50°C, during first strand synthesis

 

Contents

ComponentVolume    ExcelRT™ Reverse Transcriptase (200 U/μl) 100 μl      RNase Inhibitor (20 U/μl)100 μl      5X RT Buffer (DTT)500 μl      dNTPs Mix (10 mM each)200 μl      Oligo (dT)20 (50 μM)100 μl      Random Hexamers (100 μM)100 μl      DEPC-Treated H2O  1 ml x 2

Storage Buffer

Reverse Transcriptase: 20 mM Tris-HCl (pH 7.5), 200 mM NaCl, 0.1 mM EDTA, 1 mM DTT, stabilizer and 50% (v/v) glycerol

RNase Inhibitor: 40 mM HEPES-KOH (pH 7.5), 100 mM KCl, 8 mM DTT, 0.1 mM EDTA, stabilizer and 50% (v/v) glycerol

5X RT buffer (DTT)

250 mM Tris-HCl (pH 8.3), 375 mM KCl, 15 mM MgCl₂ and 50 mM DTT

Storage

-20°C for 24 months 

ExcelRT™ Reverse Transcription Kit is a complete, efficient and convenient kit to synthesize high quality first strand cDNA. This kit contains ExcelRT™ Reverse Transcriptase, which is able to synthesize the first strand cDNA at 37~50°C. The ExcelRT™ Reverse Transcriptase is a recombinant Moloney Murine Leukemia Virus (M-MLV) reverse transcriptase, which is designed to reduce RNase H activity and create better thermal stability. This kit also contains RNAok™ RNase Inhibitor, which is active against RNase A, RNase B, and RNase C. This product is supplied with oligo (dT)20 and random hexamers, which are used to synthesize cDNA from poly(A) tailed mRNA and total RNA, respectively.