Description 

The ExcelTaq™ 5X/2X PCR Master Mix is a ready-to-use mixture for amplifying targeted DNA fragments. It is designed to serve as a master mix for virtually all PCR applications. The mixture contains all components for PCR with the exception of templates and primers. This not only saves valuable time in the laboratory, but also reduces pipetting and reagent handling errors. The PCR Master Mix is supplied as a 5X/2X concentrated ready-to-use mixture of recombinant Taq DNA Polymerase, reaction buffer, MgCl₂ (TP1120 contains MgSO4), dNTP, and enzyme stabilizer enabling efficient amplification of template in PCR and allows the user to prepare a PCR reagent conveniently. This product is supplied with 6X DNA Loading Dye (Blue) containing two tracking dyes (Xylene cyanol FF and Bromophenol blue) for post PCR analysis through the use of agarose gel electrophoresis.

Features

• 5’→3’ DNA polymerase activity
• No detectable 3’→5′ exonuclease (proofreading) activity
• Generates PCR products with 3′-dA overhangs
• High yield PCR 
• High reproducibility
• Reduced pipetting errors

Applications 

• Routine PCR
• Colony PCR
• High throughput PCR
• Amplification of DNA fragments up to 8 kb
• Generation of PCR products for TA cloning
• DNA labeling

Storage

4°C for 6 months
-20°C for 24 months

The ExcelTaq™ 5X/2X PCR Master Mix is a ready-to-use mixture for amplifying targeted DNA fragments. It is designed to serve as a master mix for virtually all PCR applications. The mixture contains all components for PCR with the exception of templates and primers. This not only saves valuable time in the laboratory, but also reduces pipetting and reagent handling errors. The PCR Master Mix is supplied as a 5X/2X concentrated ready-to-use mixture of recombinant Taq DNA Polymerase, reaction buffer, MgCl2 (TP1120 contains MgSO4), dNTP, and enzyme stabilizer enabling efficient amplification of template in PCR and allows the user to prepare a PCR reagent conveniently. This product is supplied with 6X DNA Loading Dye (Blue) containing two tracking dyes (Xylene cyanol FF and Bromophenol blue) for post PCR analysis through the use of agarose gel electrophoresis.

Introduction

HiPure Gel Pure DNA Kit uses proprietary chemistry and HiPure technology to recover DNA fragments between 60bp-10kbp with yields exceeding 80%. DNA is suitable for ligations, PCR, sequencing, restriction digestion, or various labeling reactions. In addition, this kit can be also used to recover DNA directly from enzymatic reactions such as PCR and enzyme digestion reactions.

Details

Specifications

Features	Specifications
Main Functions	Recover DNA fragments >100bp from agarose gel(<0.5g), purification of DNA from PCR, enzymatic reaction solution or crude gDNA
Applications	PCR, NGS, labeling, ligation and enzyme digestion, etc.
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	Agarose gel, PCR products, enzyme products
Sample amount	Agarose gel: ≤500mg
Recovery	≥80%
Elution volume	≥15μl
Time per run	≤20 minutes（1-24 samples）
Liquid carrying volume per column	800µl
Binding yield of column	35µg

Principle

The HiPure system uses a simple bind-wash-elute procedure. Gel slices are dissolved in a buffer containing a pH indicator, allowing easy determination of the optimal pH for DNA binding, and the mixture is applied to the column. Nucleic acids adsorb to the silica-gel membrane in the high-salt conditions provided by the buffer. Impurities are washed away and pure DNA is eluted with a small volume of low-salt buffer provided or water, ready to use in subsequent applications.

Advantages

• High recovery efficiency – ≥80% DNA recovery
• General – recover DNA from gel or enzyme-driven reaction solutions such as PCR
• Fast – isolation can be completed in 10-15 minutes by column gel method
• Great cost-effectiveness performance

Kit Contents

Contents	D211102	D211103
Purification Times	100 Preps	250 Preps
Buffer GDP	120 ml	250 ml
Buffer DW2	50 ml	2 x 50 ml
Elution Buffer	20 ml	30 ml
HiPure DNA Mini Columns II	100	250
2 ml Collection Tubes	100	250

Storage and Stability

The Kit should be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions. If any precipitates form in the buffers, warm at 37℃ to dissolve.

Experiment Data

HiPure Gel Pure DNA Kit uses proprietary chemistry and HiPure technology to recover DNA fragments between 60bp-10kbp with yields exceeding 80%. DNA is suitable for ligations, PCR, sequencing, restriction digestion, or various labeling reactions. In addition, this kit can be also used to recover DNA directly from enzymatic reactions such as PCR and enzyme digestion reactions.

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