K-MBG4

SKU: 700004319

100 assays (manual) / 400 assays (auto-analyser)

Content:	(Malt β-glucanase) 100 assays (manual) / 400 (auto-analyser) Or (Lichenase) 100 / 200 assays (manual) / 330 (auto-analyser)
Shipping Temperature:	Ambient
Storage Temperature:	Short term stability: 2-8oC, Long term stability: See individual component labels
Stability:	> 2 years under recommended storage conditions

Analyte:	β-Glucanase/Lichenase
Assay Format:	Spectrophotometer, Auto-analyser
Detection Method:	Absorbance
Wavelength (nm):	400
Signal Response:	Increase
Limit of Detection:	(Malt β-glucanase) 4.3 x 10-4 U/mL Or (Lichenase) 9.1 x 10-5 U/mL
Reproducibility (%):	~ 3%
Total Assay Time:	(Malt β-glucanase) ~ 20 min Or (Lichenase) ~ 10 min
Application examples:	Crude malt extracts, industrial enzyme preparations.
Method recognition:	Novel method

The MBG4 reagent contains a single substrate, namely 4,6-O-benzylidene-2-chloro-4-nitrophenyl-β-(3¹-β-D-cellotriosyl-glucoside) (BCNPBG4). The benzylidene acetal group prevents any hydrolytic action by exo-acting hydrolytic enzymes such as β-glucosidase or cellobiohydrolase.

Mixed linkage β-glucanase (endo-1,3:1,4-β-glucanase) / lichenase (EC 3.2.1.73) acts specifically to release 2-chloro-4-nitrophenol (CNP) from this substrate. The rate of release of CNP is directly related to the β-glucanase/lichenase activity in a sample. The reaction is terminated and the phenolate colour is developed on addition of Tris buffer solution (pH = 10.0).

Note that the substrate is not hydrolysed by β-glucosidase or cellobiohydrolase. The substrate can be hydrolysed by certain endo-cellulases (e.g. Trichoderma sp.) but this does not result in an increase in absorbance.

Discover more assay kits for enzyme activity measurement.

Data calculators are located in the Documents tab.

Advantages

• Very cost effective 
• All reagents stable for > 2 years 
• Specific for endo-1,3:1,4-β-glucanase/lichenase 
• Simple, convenient, rapid assay 
• Well suited to automation 
• Malt flour standard and lichenase standard included

The MBG4 reagent contains a single substrate, namely 4,6-O-benzylidene-2-chloro-4-nitrophenyl-β-(31-β-D-cellotriosyl-glucoside) (BCNPBG4). The benzylidene acetal group prevents any hydrolytic action by exo-acting hydrolytic enzymes such as β-glucosidase or cellobiohydrolase.

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

Soil samples contain a large number of microorganisms, the vast majority of which can not be directly cultivated for reproduction and research. Extracting DNA from soil samples is the most effective method for studying soil microorganisms. At present, there are mainly direct and indirect methods for extracting microbial DNA from soil samples. The direct method refers to placing soil samples in the lysis solution, and using effective wall breaking methods to release all microbial DNA into the lysis solution, followed by separation and extraction, such as Zhou’s method. Indirect method refers to placing soil in a buffer, such as Buffer PBS, to separate microorganisms from the soil and then extract DNA. The indirect method can greatly reduce the impact of humic acids and heavy metal salts on DNA extraction in soil, but this method will lose many microorganisms and the resulting DNA is not the entire genome (metagenome) of the soil sample. Currently, few researchers have adopted this method. Extracting DNA directly from soil samples can maximize the likelihood of obtaining the entire genome, but this method faces the following issues:

1. Humic acid pollution. The soil, especially in forests and grasslands, is rich in humic acids. Humic acid is a series of organic molecules, some of which are very similar to nucleic acid molecules and difficult to remove during purification. Trace amounts of humic acid pollution can lead to downstream applications such as PCR and enzyme digestion failure.

2. Lysis method. Soil samples contain various microorganisms, such as bacteria and fungi. Gram positive bacteria and fungi both contain very thick bacterial walls, and effectively breaking down the cell walls of these microorganisms is crucial for extracting high-yield metagenomic DNA. Due to the complexity of soil samples, it is not feasible to use enzymatic methods (such as lysozyme, wall breaking enzyme, snail enzyme) or liquid nitrogen grinding, as the soil contains various metalions or inhibitory factors that inactive the digestive enzymes, or the presence of sand particles in the soil makes liquid nitrogen grinding difficult.

3. The DNA yield is difficult to control. Soil samples would have significant changes in the number and variety of microorganisms due to fertility, inferiority, high moisture content, dryness, or depth of sampling. In a small range of soil samples, the DNA content often varies by thousands of times. In addition, certain chemical components in soil, such as heavy metal salts and clay substances, can cause a decrease in DNA yield.

Magen’s HiPure Soil DNA Kits are currently the most optimized kit for soil DNA extraction. The kit adopts glass bead grinding method and thermal shock chemical wall breaking method, which can be carried out in the point vortex instrument without special bead grinding instrument, and is suitable for a wide range of laboratories. The Absorber Solution in the reagent kit is a humic acid adsorbent exclusively developed by Magen Company, which can efficiently remove various humic acid pollutants. In addition, an alcohol-free silica gel column purification method is also used to efficiently remove various soluble metal salts and other soluble inhibitory factors from the soil. The kit has successfully extracted from the following soil (partially based on customer feedback): soil from forests in nature reserves (30 to 40 years old forest soil with a surface layer of 30-50cm deciduous layer), mangrove soil, grasslands, farmland, seabed mud, sludge, mineral area soil, organic matter contaminated soil, pond mud, garbage mud, air conditioning pipeline deposits, etc.

This product allows rapid and reliable isolation of high-quality genomic DNA from various soil samples. Up to 500 mg soil samples can be processed in 60 minute. The system combines the reversible nucleic acid binding properties of HiPure matrix with the speed and versatilityof spin column technology to eliminate PCR inhibiting compounds such as humic acid from soil samples. Purified DNA is suitable for PCR, restriction digestion, and next-generation sequencing. There are no organic extractions thus reducing plastic waste and hands-on time to allow multiple samples to be processed in parallel.

Details

Specifications

Features	Specifications
Main Functions	Isolation DNA from 200-500mg soil sample
Applications	PCR, southern blot and enzyme digestion, etc.
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	Soil
Sample amount	200-500mg
Elution volume	≥30μl
Time per run	≤60 minutes
Liquid carrying volume per column	800μl
Binding yield of column	100μg

Principle

Soil sample is homogenized and then treated in a specially formulated buffer containing detergent to lyse bacteria, yeast, and fungal samples. humic acid,proteins, polysaccharides, and other contaminants are removed using our proprietary Absorber Solution. Binding conditions are then adjusted and the sample is applied to a DNA Mini Column. Two rapid wash steps remove trace contaminants and pure DNA is eluted in low ionic strength buffer. Purified DNA can be directly used in downstream applications without the need for further purification.

Advantages

• Fast – several samples can be extracted in 40 minutes (after digestion)
• High purity – purified DNA can be directly used in various downstream applications
• Good repeatability – silica technology can obtain ideal results every time
• High recovery – DNA can be recovered at the level of PG

Kit Contents

Contents	D314202	D314203
Purification Times	50 Preps	250 Preps
Hipure DNA Mini Columns II	50	250
2ml Collection Tubes	50	250
2ml Bead Tubes	50	250
Buffer SOL	60 ml	250 ml
Buffer SDS	5 ml	20 ml
Buffer PS	10 ml	50 ml
Absorber Solution	10 ml	50 ml
Buffer GWP	40 ml	220  ml
Buffer DW1	30 ml	150  ml
Buffer GW2*	20 ml	2 x 50  ml
Buffer AE	15 ml	30 ml

Storage and Stability

Absorber Solution 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.

Experiment Data

Soil samples contain a large number of microorganisms, the vast majority of which can not be directly cultivated for reproduction and research. Extracting DNA from soil samples is the most effective method for studying soil microorganisms. At present, there are mainly direct and indirect methods for extracting microbial DNA from soil samples. The direct method refers to placing soil samples in the lysis solution, and using effective wall breaking methods to release all microbial DNA into the lysis solution, followed by separation and extraction, such as Zhou’s method. Indirect method refers to placing soil in a buffer, such as Buffer PBS, to separate microorganisms from the soil and then extract DNA. The indirect method can greatly reduce the impact of humic acids and heavy metal salts on DNA extraction in soil, but this method will lose many microorganisms and the resulting DNA is not the entire genome (metagenome) of the soil sample. Currently, few researchers have adopted this method. Extracting DNA directly from soil samples can maximize the likelihood of obtaining the entire genome, but this method faces the following issues: