Introduction

This product provides fast and easy methods for purification of total DNA for reliable PCR and Southern blotting. Total DNA (e.g., genomic, viral, mitochondrial) can be purified from tissue and culture cells.

Details

Specifications

This product is based on silica Column purification. The sample is lysed and digested with lysate and protease, DNA is released into the lysate. Transfer to an adsorption column. Nucleic acid is adsorbed on the membrane, while proteinis not adsorbed and is removed with filtration. After washing proteins andother impurities, Nucleic acid was finally eluted with low-salt buffer (10mmTris, pH9.0, 0.5mm EDTA).

Advantages

• Good repeatability – suitable for extracting high-yield DNA from different types of tissue samples
• High purity – can be used for downstream applications such as multiplex and quantitative PCR
• Fast – simplified process, extracting several samples in 20 minutes (after digestion)
• Safety – no phenol orchloroform extraction, no alcohol precipitation

Kit Contents

Storage and Stability

RNase A and Proteinase K 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 its 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. The entire kit can be stored at 2-8°C, but in this case buffers should be redissolved before use. Make sure that all buffers are at room temperature when used.

Purchase Guide

This product provides fast and easy methods for purification of total DNA for reliable PCR and Southern blotting. Total DNA (e.g., genomic, viral, mitochondrial) can be purified from tissue and culture cells.

K-MRGLUC

1000 assays (microplate) / 100 assays (manual) / 400 assays (auto-analyser)

This product has been discontinued.

The D-Glucose Assay Kit (Megaplex Red) provides a simple robust method for the measurement of D-glucose. This kit utilises a highly sensitive Megaplex Red probe which when coupled to horseradish peroxidase (HRP) and glucose oxidase (GOX) enzymatic reactions, allows for the measurement of D-glucose in a sample. Upon oxidation of the probe by HRP, a coloured product (resorufin) is formed that can be measured in fluorescence mode using excitation between 530-560 nm and emission at ~ 590 nm* or colourimetrically at 570 nm.

*The excitation and emission maxima of Megaplex Red are 570 nm and 585 nm respectively.

See more related mono/disaccharide assay kit products. 

Fluorometric/UV method for the determination of D-glucose content in a variety of samples.

Advantages

• Simple format
• Very competitive price (cost per test)
• Ability to run in fluorescence or absorbance mode
• User friendly – Detailed protocol provided for the creation of calibration curve and the calculation of concentration in fluorescence mode. Single point standard for quicker and simpler analysis in absorbance mode.
• Mega-Calc™ software tool is available from our website for hassle-free raw data processing
• Suitable for manual, microplate and auto-analyser formats
• All reagents stable for > 2 years

The D-Glucose Assay Kit (Megaplex Red) provides a simple robust method for the measurement of D-glucose. This kit utilises a highly sensitive Megaplex Red probe which when coupled to horseradish peroxidase (HRP) and glucose oxidase (GOX) enzymatic reactions, allows for the measurement of D-glucose in a sample. Upon oxidation of the probe by HRP, a coloured product (resorufin) is formed that can be measured in fluorescence mode using excitation between 530-560 nm and emission at ~ 590 nm* or colourimetrically at 570 nm.

Introduction

HiPure RNA Clean Up Kit provides a rapid and easy method for the purification and concentrate RNA from enzymatic reactions or for desalting the RNA samples. RNA purified using HiPure RNA Clean Up Kit is ready for all downstream applications such as RT-PCR, Northern blotting, mRNA purification, nuclease protection, and in vitro translation.

Details

Specifications

Principle

The HiPure system uses a simple bind-wash-elute procedure. Binding buffer is added directly to the sample or other enzymatic reaction, and the mixture is applied to the column. Nucleic acids adsorb to the silica membrane in the high-salt conditions provided by the buffer. Impurities are washed away and pure RNA is eluted with a small volume of low-salt buffer provided or water, ready to use in all subsequent applications.

Advantages

• High recovery – recovery of RNA up to 90%
• Low elution volume – the elution volume can be as low as 15µl
• Fast – only 10 minutes for recovery by using column method
• General – suitable for various crude RNA products
• Wide range of fragment recovery:(>25nt RNA)

Kit Contents

Storage and Stability

The Kit can be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions. The entire kit can be stored at 2-8°C, but in this case buffers should be redissolved before use. Make sure that all buffers are at room temperature when used.

HiPure RNA Clean Up Kit provides a rapid and easy method for the purification and concentrate RNA from enzymatic reactions or for desalting the RNA samples. RNA purified using HiPure RNA Clean Up Kit is ready for all downstream applications such as RT-PCR, Northern blotting, mRNA purification, nuclease protection, and in vitro translation.

Endonucleases Non-Specific, HL-SAN

OverView

HL-SAN efficiently removes nucleic acids from buffers typically used in protein purification. Due to its high salt tolerance, it is the obvious choice for host-cell DNA removal in settings where salt is added to reduce aggregation. Especially efficient for removing nucleic acids from proteins with high affinity for DNA and RNA. Proven performance during lysis and early stages of protein purification processes, as well as high-salt eluates. Cold-adapted enzyme with excellent performance also at ambient temperatures and during over-night digestion at 4°C.

Recommended applications:

Complete removal of DNA 

Viscosity reduction

Key advantages with this Salt Active Nuclease:

• Non specific endonuclease
• Optimum activity at high salt concentration (0.5 M NaCl)
• Active at low temperatures (20% at 6ºC)
• Easily inactivated
• Broad pH range
• Temperature stable

Figures

Figure 1. Optimum activity in solutions with high salinity

HL-SAN has optimum activity at ∼0.5 M NaCl, but operates at a broad range of [NaCl] and [KCl]. The activity of HL-SAN was tested in a 25 mM Tris-HCl buffer, pH 8.5, 5 mM MgCl₂ with varying [NaCl] or [KCl]. The maximum activity was set to 100%.

Figure 2. Temperature and activity

HL-SAN has optimum activity at ~35°C, but works over a broad temperature range (20% activity at 10°C and 50°C). The activity of HL-SAN was tested in a 25 mM Tris-HCl buffer, pH 8.5 containing 5 mM MgCl₂ and 0.5 M NaCl.

Fig 3. The effect of MgCl₂ and MnCl₂ concentration on the HL-SAN activity.

The activity of HL-SAN was tested in a 25 mM Tris-HCl buffer, pH 8.5, 0.5 M NaCl and with varying concentrations of MgCl₂ or MnCl₂. The activity of the sample containing 5 mM MgCl₂ was set to 100%.

Figure 4. HL-SAN activity vs pH/[NaCl]

The activity of HL-SAN was tested in a 25 mM Tris-HCl buffer with different pHs and different concentrations of NaCl. All buffers contained 5 mM MgCl₂. The nature of the buffer was pH-dependent, but generally the NaCl-optimum was the same in all buffers/pHs. The exception was etanolaminbuffer at pH 9 and pH 9.5 in which the NaCl-optimum was shifted to the left (not shown).

Figure 5. Buffer composition affects substrate preference

Without NaCl, the specificity towards ssDNA and dsDNA is similar. At 0.5 M NaCl, the activity towards dsDNA increases, while the activity towards ssDNA is unaffected.

Figure 6. HL-SAN digests ssDNA to ~5-13 nt, and dsDNA to ~5-7 nt

The size of the end products from ssDNA varies from ~5-13 nt, while dsDNA is digested to around ~5-7 nt. The size of the end products seems to depend on the DNA sequence. Substrates 1 and 2 were ssDNA with different sequences and substrates 3 and 4 were dsDNA with similar sequences but with a FAM-label at different ends. Substrate 5 was dsDNA with the same sequence as substrate 3 and 4 but with a FAM-label at both ends.

‍

Figure 7. HL-SAN activity decreases with increasing concentrations of glycerol

The activity of HL-SAN was tested in a 25 mM Tris-HCl buffer, pH 8.5, 5 mM MgCl₂, 0.5 M NaCl and with increasing concentrations of glycerol. The activity of the control not containing glycerol was set to 100%.

Figure 8. The activity of HL-SAN at different concentrations of imidazole

The activity of HL-SAN was tested in a 25 mM Tris-HCl buffer, pH 8.5, 5 mM MgCl₂, 0.5 M NaCl and with varying concentrations of imidazole. The activity of the control not containing imidazole was set to 100%.

HL-SAN efficiently removes nucleic acids from buffers typically used in protein purification. Due to its high salt tolerance, it is the obvious choice for host-cell DNA removal in settings where salt is added to reduce aggregation. Especially efficient for removing nucleic acids from proteins with high affinity for DNA and RNA. Proven performance during lysis and early stages of protein purification processes, as well as high-salt eluates. Cold-adapted enzyme with excellent performance also at ambient temperatures and during over-night digestion at 4°C.

K-GLUC

SKU: 700004297

660 assays per kit

The D-Glucose test kit contains high purity reagents for the measurement and analysis of D-glucose in cereal extracts and for use in combination with other Megazyme kits.

See more related mono/disaccharide assay kit products.

Advantages

• All reagents stable for > 12 months after preparation 
• Very competitive price (cost per test) 
• Simple format 
• Standard included

The D-Glucose test kit contains high purity reagents for the measurement and analysis of D-glucose in cereal extracts and for use in combination with other Megazyme kits.