NucleoBond HMW DNA
Short-read NGS technologies have dramatically accelerated the pace of biological discovery in recent years, but for applications such as genome assembly, analysis of repetitive regions, and genome phasing, longer-read approaches are often a better fit. The NucleoBond HMW DNA kit was specifically developed for long-read sequencing applications, and employs gravity flow filtration in combination with NucleoBond anion-exchange technology to enable efficient recovery of high molecular weight genomic DNA of the highest purity and integrity from diverse samples.
Short-read NGS technologies have dramatically accelerated the pace of biological discovery in recent years, but for applications such as genome assembly, analysis of repetitive regions, and genome phasing, longer-read approaches are often a better fit. The NucleoBond HMW DNA kit was specifically developed for long-read sequencing applications, and employs gravity flow filtration in combination with NucleoBond anion-exchange technology to enable efficient recovery of high molecular weight genomic DNA of the highest purity and integrity from diverse samples.
NucleoBond HMW DNA provides a gentler alternative to magnetic bead-based approaches, which include extensive pipetting and mixing steps that threaten DNA integrity. It also provides superior performance vs. liquid phase methods (e.g., salting out), which can be tedious and time-consuming and tend to yield DNA of lower purity. The kit enables isolation of high-quality DNA up to 200 kb in length from a wide variety of starting materials—including gram-positive and gram-negative bacteria, yeast, plants, insects, mammalian tissue, blood, and cultured cells—and is designed to accommodate large input amounts (e.g., up to 1.5 g of plant material or up to 300 mg of mammalian tissue per prep). The versatile kit protocol enables processing of 12 samples in about two hours and can be combined with additional upstream and downstream processing steps, including enzymatic or mechanical sample lysis (e.g., MN bead tubes) and DNA concentration/desalinization (e.g., NucleoSpin and NucleoSnap finishers). DNA purified with the kit is ideal for analysis by long-read sequencing technologies such as SMRT (PacBio) and Nanopore (Oxford Nanopore), as well as long-range short-read methods such as Next GEM (10X Genomics).
Overview
- High quality, high molecular weight (HMW) DNA up to 200 kb
- Efficient processing of large input amounts
- Minimized DNA shearing due to established anion-exchange technology
- Validated with diverse samples and sequencing platforms
More Information
Technology | Anion-exchange chromatography |
Format | Midi gravity flow columns |
Fragment size | Up to 200 kb with >90% depletion of fragments <10 kb |
Sample material |
<1.5 g plant leaves (ground under liquid nitrogen) <107 cultured cells (enzymatic lysis) <300 mg solid tissue (cut into small pieces and lysed enzymatically with increased lysis time, ground under liquid nitrogen, or lysed by bead beating) <30 mg yeast or bacteria (ground under liquid nitrogen) <300 mg yeast or bacteria (lysed enzymatically or by bead beating) <2 ml liquid sample (e.g., blood, body fluids, or enzymatic reactions) |
Typical yield | Depends on sample type and amount |
Elution volume | 50–250 µl |
Processing time | 2 hr/12 preps (including 30-min lysis) |
Applications
- Isolation of high molecular weight DNA from mammalian tissues, blood samples, invertebrates, plants, bacteria, and yeast
- Typical downstream applications: long-read sequencing (e.g., SMRT, Nanopore, Next GEM)
Additional product information
Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.
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