Features

Magnet bead based high throughput isolation of DNA from plant samples

• Optimized for high throughput processing in a 384-well format
• Efficient purification of DNA from up to 30 mg sample material
• Suitable for a wide variety of plant species

 

Technology	Magnetic bead technology
Format	Highly reactive superparamagnetic beads
Processing	Manual or automated
Sample material	< 30 mg plant tissue (wet weight)
Plant species already successfully tested	Zea mays (maize), Brassica napus (rapeseed), Glycine max (soy), Triticum aestivum (wheat), Nicotiana tabacum (tobacco), Elaeis guineensis (oil palm*)
Typical yield	Depending on sample source, storage, and quality
A260/A280	1.6–1.9
Elution volume	40–100 µL
Preparation time**	40–120 min/96 preps (excl. lysis) 60 min/384 preps (excl. lysis)
Binding capacity	0.2 µg/µL beads

*Support protocol available. Please inquire technical support.
**Depending on instrument type, setup, and configuration.

Applications

• Manual or automated isolation of DNA from plant tissue
• Typical downstream applications: PCR, Southern blotting, any kind of enzymatic reaction

 

Principle / Procedure

The NucleoMag® 384 Plant kit enables efficient and rapid isolation of genomic DNA from a wide range of plant species and plant organs as an integral step in plant research or plant breeding laboratories. The kit composition was optimized for reliable high throughput processing of sample material up to 30 mg and is suitable for 384-well formats. The NucleoMag® 384 Plant kit was designed to deal with typical issues during plant sample processing, such as the release of polyphenolic compounds or complex polysaccharides.

Plant tissue is extracted with CTAB Lysis Buffer MC1 and binding of DNA to the NucleoMag® V-Beads is facilitated with Binding Buffer MC2. After magnetic separation and removal of supernatant, the paramagnetic beads are washed with Wash Buffers MC3, MC4, and 80 % ethanol to remove contaminants and salt. There is no need for a drying step as ethanol from previous wash steps is removed by Wash Buffer MC5. Finally, DNA is eluted with low salt Elution Buffer MC6 and is ready to use for downstream applications such as NGS, qPCR, blotting, or enzymatic reactions. The NucleoMag® 384 Plant is designed for automated high throughput processing on standard liquid handling instruments in a 384-well format.

Application data

Reliable detection of genomic DNA from various different plant species
DNA was isolated from 30 mg fresh leaves or seeds from different plant species using the NucleoMag® 384 Plant kit. The total yield was determined by UV spectrometry (dark blue bars). A subsequent qPCR analysis (orange squares) was performed with a Taqman® Probe for a 103 bp actin amplicon using the SensiFastTM Probe Lo-ROX kit from Bioline on an Applied Biosystems® 7500 Real-Time PCR System. High throughput processing with the NucleoMag® 384 Plant kit of all tested sample material, results in pure and sufficient DNA yields, ready for direct subsequent biomolecular analysis.

** Genomic DNA extraction of oil palm leaves was achieved by addition of Binding Buffer CB according to the support protocol NucleoMag® 384 Plant – Oil palm leaves.