Auto-Mag® HMW DNA Isolation Kit

Auto-Mag® HMW DNA Isolation Kit is specifically designed for isolation of high molecular weight DNA (HMW DNA) in the size range of 50–300+ kb. The kit extracts HMW DNA from biological samples such as whole blood, bone marrow, saliva, buccal cells, cultured cells, tissues, and bacteria. 

• Reproducibly purify high molecular weight genomic DNA (50-300+kb) from a wide range of samples matrices.
• Simplified, user friendly protocols that remove RNA and inhibitors.
• Comes with enzymes mixture for isolation high quality HMW DNA from Bacteria
• HMW DNA is suitable for all third-generation sequencing platforms including Nanopore and PacBio SMRT sequencing.
• Automation-friendly and can be easily scaled up.

The Auto-Mag® HMW DNA Isolation Kit utilizes magnetic bead-based technology in combination with chaotropic agents to gently isolate HMW DNA and remove inhibitors. Kit contains sufficient reagents for the specified number of preparations. The kit also includes RNase A for RNA removal and a special lytic enzyme mix for bacterial HMW DNA Isolation. The HMW DNA purified with this kit is particularly suitable for analysis on long read sequencing genomic platforms including PacBio RSII/Sequel/Sequel II and Oxford Nanopore.

The size of the extracted HMW DNA can depend on the sample matrix, the quality of the starting material, and the processing parameters. High-quality samples are critical to obtaining high-quality HMW DNA. This kit can be used with fresh or frozen samples, but care should be taken to minimize freeze-thaw cycles. To maximize the integrity of the extracted genomic DNA, avoid excessive mixing samples with pipettes during the extraction and isolation process, except for the elution step. In addition, if you encounter low-quality samples or partially degraded extracted DNA, it is recommended to further purify the extracted DNA using our unique Auto-Mag® HMW Size Select reagent (Cat. No. S005) to remove small DNA fragments from the DNA sample and recover HMW DNA of the desired size to meet the needs of long-read sequencing genomic platforms.

Auto-Mag® HMW DNA Isolation Kit uses magnetic technology and requires no centrifugation or filtration, it is suitable for manual processing as well as adapting to automatic liquid handling workstations.

Manual Workflow for HMW DNA Isolation from Blood Sample

1. Confirm the sample volume.
2. Add Proteinase K to lysis the cells.
3. Add Beads to bind DNA
4. Separate beads from contaminants.
5. Wash magnetic beads to remove contaminants.
6. Elute DNA from magnetic beads.

Data & References

HMW DNA Isolation

Fig 1. HMW gDNA  were isolated from whole blood, saliva or cultured cell samples with AMD’s Auto-Mag® HMW DNA Isolation Kit. The isolated HMW DNA was analyzed  on Agilent’s TapeStation 2200.

Comparision of HMW DNA Purity & Yield

Figure 2. HMW Genomic DNA was extracted from 200 µl of whole blood using Auto-Mag® HMW DNA Isolation Kit and Competitor (M) HMW DNA Isolation Kit. Purity and concentration of DNA extracted by both kits was assessed using NanoDrop spectrophotometer. The measurements were determined in triplicate. The ratio of absorbance at 260 nm and 280 nm was used to assess protein contamination while the ratio of absorbance at 260 nm and 230 nm was used to assess the presence of contaminants. Both kits’ spectrophotometric measurements met the criteria for high quality DNA. In terms of determined recovery, Auto-Mag® HMW DNA Isolation Kit had a slightly better DNA yield compared to Competitor (M) kit.