Description
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).
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 C T 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%
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.
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).
