MagAttract HMW DNA Kit

For isolation of high-molecular-weight genomic DNA

S_1084_5_GEN_V2

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MagAttract HMW DNA Kit (48)

Cat. No. / ID:   67563

For 48 DNA preps: MagAttract Suspension G, Buffer ATL, Buffer AL, Buffer MB, Buffer MW1, Buffer PE, Proteinase K, RNase A, Buffer AE, Nuclease-Free Water
The MagAttract HMW DNA Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

Features

  • Reproducible isolation of high-molecular-weight DNA
  • High yields and purity from a range of sample materials
  • Fast and convenient protocols
  • Complete removal of inhibitors

Product Details

The MagAttract HMW DNA Kit enables purification of high-molecular-weight (100–200 kb) DNA using a simple, magnetic bead-based protocol. The purification procedure gently removes contaminants and inhibitors, while delivering very high yields and purity. The extracted genomic DNA is suitable for numerous applications, including archiving and next-generation sequencing (NGS).

Performance

The MagAttract HMW DNA Kit ensures reproducible isolation of genomic DNA >150 kb. DNA isolated using the MagAttract HMW DNA Kit and other magnetic bead-based procedures was compared using pulse-field gel electrophoresis. The MagAttract HMW DNA Kit demonstrated superior performance in the isolation of high-molecular-weight DNA (see figure  Successful isolation of high-molecular-weight DNA).


Consistent high yields and purity
The MagAttract HMW DNA Kit uses a convenient, straightforward procedure to deliver high yields of pure, high-molecular-weight DNA. Gentle lysis conditions minimize DNA fragmentation, while contaminants and PCR inhibitors are efficiently removed, resulting in pure, high-molecular-weight DNA. The optimized chemistry and procedure of the MagAttract HMW DNA Kit greatly enhance DNA yield and purity, compared to classic spin-column or magnetic bead-based methods from other suppliers (see figures  Highest yields and purity and  Improved yields from various sample types).


Effective removal of PCR inhibitors
Highly sensitive applications such as real-time PCR or NGS require the removal of anticoagulants, enzymes, divalent cations, and other such inhibitors. Real-time PCR amplification of DNA isolated from blood samples stabilized with EDTA, heparin, and citrate stabilized using the MagAttract HMW DNA Kit was performed. No inhibition is seen in any of the samples, which demonstrate highly linear correlation of CT values with sample volume input (see figure  No inhibition in samples derived from anticoagulated blood).


Genomic DNA ready for next-generation sequencing
Since massively parallel sequencing (MPS) technologies utilize short read lengths, sequencing can be performed using intact genomic DNA >10 kb. De novo genome assembly and the discovery of inherited and acquired structural variants are, however, still challenging with standard DNA purification products. Mate-pair libraries with larger inserts may have significant impact on sequencing success, especially for complex repeat-rich genomes. The high-molecular-weight genomic DNA obtained with the MagAttract HMW DNA Kit is ready for use in NGS experiments

Representative results of a next-generation sequencing experiment (mate-pair)
Parameter Specification
Total (long) mapped reads 6521106
Error rate 0.36%
Indel rate 0.01%
Chimeras 0.14%
Percent duplication 1.15%
See figures

Principle

Isolation of genomic DNA is a crucial step for any DNA analysis method. The key parameters are generally yield, purity, and integrity of the isolated DNA. While for standard methods such as PCR, sequencing, or hybridization techniques, molecular weight in the range of 10–30 kb may be sufficient, some sensitive methods such as the generation of BAC or fosmid libraries or archiving of DNA require higher integrity of genomic DNA.
The MagAttract HMW DNA Kit is specifically designed to deliver high-molecular-weight DNA from a wide range of samples, including human or animal tissue, blood, and bacteria. The kit uses an efficient combination of high-performance magnetic beads and innovative silica-based chemistry. High-performance magnetic particles ensure that the isolated DNA is of high yield and purity.

Procedure

The convenient MagAttract protocol allows reproducible results in as little as 70 minutes.


Fast and optimized protocol
Optimized buffers and enzymes gently lyse samples, while ensuring minimized fragmentation of genomic DNA. The procedure comprises 4 simple steps: lyse, bind, wash, and elute. Following sample lysis, the DNA binds to the surface of magnetic beads. During the wash steps, contaminants and PCR inhibitors are effectively removed and pure, high-molecular-weight DNA is eluted in Buffer AE (see figure  MagAttract HMW DNA Kit procedure).

See figures

Applications

The high-molecular-weight genomic DNA obtained with the MagAttract HMW DNA Kit is ready for use in NGS experiments. It can be used for the generation of libraries with both small and large inserts and is therefore suitable for all sequencing applications.

Supporting data and figures

Specifications

FeaturesSpecifications
Sample amount200 μl blood; up to 25 mg tissue; 2 x 10e9 bacterial cells
Process time~70 minutes for 12 samples
Elution volume100–200 µl

Resources

Brochures & Guides (1)
Introducing QIAseq
PDF (450KB)
Accelerate your NGS performance through Sample to Insight solutions
Kit Handbooks (1)
For isolation of high-molecular-weight DNA from blood, tissue, and bacteria for next-generation sequencing applications
Quick-Start Protocols (1)
Safety Data Sheets (1)
Certificates of Analysis (1)

FAQ

Can I scale up reagents if I want to use more sample with the MagAttract HMW DNA Kit (cat. no. 67563)?

Technically yes, however you would need to use different containers like 15 ml tubes and suitable magnets. You cannot use 2 ml tubes as the protocol states as higher amounts of beads also require more buffer for the protocol to work.

FAQ ID - 3448