High Molecular Weight DNA Isolation from Plant Tissue for Next-Generation Mapping Applications

High-quality high molecular weight (HMW) DNA is key to successful next-generation genome mapping (NGM) and other technologies that rely on HMW-DNA to conserve long range genomic information. Isolation of HMW DNA from plant tissue is especially challenging due to the tough cell wall and abundant secondary metabolites that can interfere with DNA quality. Liquid nitrogen grinding methods with homebrew reagents, traditionally used to isolate HMW DNA from plant species for BAC/BIBAC library construction, usually require 20-30 grams of starting material and often results in variable quality DNA. We have developed a simple and efficient modular workflow employing two alternate homogenization methods and three nuclei isolation strategies, which enables the isolation of high-quality HMW DNA from a wide variety of plant tissues.

The IrysPrep^® Plant Tissue DNA Isolation Kit provides essential reagents to break the plant cell wall, lyse chloroplast and mitochondria, and remove/neutralize secondary metabolites such as polyphenols and polysaccharides, while preserving/stabilizing nuclei. Microgram scale yield of HMW DNA from as little as 100 mg of fresh young plant tissue has been recovered from cotton, soybean, strawberry, blueberry, Eucalyptus, grapevine, peanut, oak, wheat, oat, tomato, maize, sunflower, brassica, coffee and successfully used for NGM.