DNeasy Plant Pro Kit
For isolation of genomic DNA from plant cells, tissues and seeds
Isolation of high-quality total DNA from plant samples is notoriously difficult due to the large number of PCR inhibitors, including polysaccharides, polyphenolics, and other secondary metabolites from plant extracts. The DNeasy Plant Pro Kit has a number of innovative features to overcome these challenges and enable recovery of high-quality DNA from the toughest sample types, including strawberry leaf, grapevine leaf, pine needles and various seed types. Increased yields of plant DNA and plant pathogen DNA in combination with superior removal of inhibitors ensure high-performance results in sensitive downstream applications.
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The DNeasy Plant Pro Kit – the newest member of the trusted DNeasy Plant family – enables purification of significantly higher yields of DNA from the toughest sample types, including strawberry leaf, grapevine leaf, pine needles and various seed types (see figure “Significantly higher yields of pure DNA”). Patented Inhibitor Removal Technology (IRT) provides superior removal of inhibitors without using harsh chemicals, yielding pure DNA with less PCR inhibition (see figure “Efficient removal of PCR inhibitors”).
When CT values of PCR reactions with plant DNA eluates containing possible inhibitors were compared to CT values of the PCR reaction with water added as control which does not inhibit amplification of the IC DNA, the eluate from the DNeasy Plant Pro Kit showed no inhibition. Using the DNeasy Plant Pro Kit, DNA was purified from strawberry leaf – a particularly tough plant sample – in direct comparison with kits from two other suppliers. Yields with the DNeasy Plant Pro Kit were significantly higher than those obtained using a kit from Supplier T and Supplier Z. Furthermore, the DNeasy Plant Pro Kit provided superior removal of inhibitors and the DNA eluate showed no inhibition (see figure “Significantly higher yields and superior inhibitor removal”). The DNeasy Plant Pro Kit can also be used to purify bacterial, fungal and viral DNA from plant and root samples.
The kit can be successfully combined into a workflow with proven products optimized for next-generation sequencing (NGS) (see figure “Optimized NGS workflow”). Rapid and reliable identification of plant pathogens is crucial to avoid disease spread and to facilitate effective disease management. Plant and pathogen DNA co-purified using the DNeasy Plant Pro Kit enables successful identification of a range of pathogens (see figure “Successful identification of plant pathogens by NGS”).
The DNeasy Plant Pro Kit comprises a novel and proprietary method for fast and easy purification of total cellular DNA from plant cells, tissues and seeds. In addition, this kit can be used to purify bacterial, fungal and viral DNA from plant and root samples. The DNeasy Plant Pro Kit uses bead-beating technology, which replaces cumbersome DNA isolation procedures such as CTAB, phenol or chloroform extraction for the recovery of high-quality DNA from the toughest sample types, including strawberry leaf, grapevine leaf, pine needles and various seed types. The DNeasy Plant Pro Kit uses the second generation of QIAGEN’s patented Inhibitor Removal Technology (IRT) to remove PCR inhibitors, including polysaccharides, polyphenolics, and other secondary metabolites from plant extracts during the isolation process. Improved IRT, in combination with efficient bead beating and lysis chemistry, results in high yields of inhibitor-free DNA that is ready for immediate use in downstream applications, including PCR, qPCR, RAPD analysis, RFLP analysis, Southern blotting and next-generation sequencing.
Samples are added to the Tissue Disruption Tube which contains a specially shaped bead and a buffer for rapid homogenization (see figure "Rapid homogenization in Tissue Disruption Tubes"). Cell lysis and DNA release occur by mechanical and chemical methods. Released genomic DNA is cleared of PCR inhibitors using QIAGEN’s second generation Inhibitor Removal Technology (IRT) and then captured on a silica membrane in a spin column format. DNA is then washed and eluted from the membrane and is ready for PCR, qPCR, NGS and other downstream applications.
An optional Phenolic Separation Solution step helps provide pure DNA even from samples with high polyphenolic compounds, such as pine or grape leaves by breaking the bonds between DNA and phenolics. This step prevents DNA loss that would otherwise occur in these samples.
The DNeasy Plant Pro Kit is used with a vortexer or high-velocity bead beating, based on sample needs. Vortex methods work with soft leaf tissue, while high-velocity bead beaters, like the PowerLyzer 24 Homogenizer or the TissueLyser II, break down tougher plant material such as roots, seeds, stems or difficult leaf tissues. Furthermore, purification of DNA using the DNeasy Plant Pro Kit can be fully automated on the QIAcube.
The DNeasy Plant Pro Kit is designed for:
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