The ClearYield DNA Isolation Kit, Plant (Item: 9K-1001P) uses a proprietary method for rapid isolation of DNA from plant samples. The DNA can be used in downstream enzymatic applications such as amplification and sequencing. The novel chemistry used in this kit replaces more laborious methods such as phenol – chloroform DNA extraction of nucleic acids. Each lysis tube contains a grinding matrix that acts in conjunction with the Lysis Buffer to release DNA and capture contaminants from the sample. Experimental data shows that the DNA Extraction Kit yields DNA of equivalent or better purity when compared to other commercial kits.
Isolation of DNA from plant samples can be challenging due to the presence of a cell wall as well as compounds such as polysaccharides, tannins, and phenolics. The Plant DNA Isolation Kit is comprised of three basic steps for the isolation of DNA from plant samples: 1) Mechanical disruption of the plant cell wall using a grinding matrix, 2) precipitation of polysaccharides and other cellular debris in the presence of a cationic detergent, and 3) isolation of DNA by RNase A treatment and purification through silica spin columns.
Fresh, frozen or lyophilized plant samples are subjected to mechanical disruption by homogenization in the presence of a grinding matrix and the Lysis Buffer. Unlike more traditional extraction methods, this method disrupts the cell wall releasing the nucleic acids into the supernatant while precipitating the cellular debris. The extracted nucleic acids contained in the supernatant are then subjected to RNase A treatment to isolate DNA that is purified using silica spin columns. The procedure takes approximately 40 minutes to complete. The purified DNA generally generates 260/280 absorbance ratios ≥ 1.8 and can be used immediately or frozen at -20 to -80oC for subsequent analysis.