Product No. E5038
Norgen’s Plant/Fungi DNA Isolation Kit provides a rapid method for the isolation and purification of total DNA from a wide range of plant and fungi species. Furthermore, the kit also provides a convenient method for the detection of pathogens which may be infecting a plant, as it allows for the purification of any pathogen DNA along with the purification of the total DNA. Total DNA can be purified from fresh or frozen plant tissues, plant cells or fungi samples using this kit. The DNA is preferentially purified from other cellular components, such as proteins, without the use of phenol or chloroform. The purified DNA is of the highest integrity, and can be used in a number of downstream applications including real time PCR, Southern blotting, SNP analysis and sequencing.
Purification is based on spin column chromatography. The DNA is preferentially purified from other cellular components such as proteins without the use of phenol or chloroform. The process involves grinding plant tissue in a mortar with liquid nitrogen (or alternative homogenization equipment). Lysis Solution and RNase A are then added, followed by a short incubation at 65 °C. Next, Binding Solution is then added to the lysate followed by another short incubation on ice. The lysate is then spun through the provided Filter Column in order to remove any debris. Ethanol is then added to the clarified lysate, and the solution is loaded onto a spin-column. Norgen’s resin binds nucleic acids in a manner that depends on ionic concentrations, thus only the DNA will bind to the column while most of the RNA and proteins are removed in the flowthrough. The bound DNA is then washed with the provided Wash Solution I and II in order to remove any remaining impurities, and the purified total DNA is eluted with the Elution Buffer. The purified DNA is of the highest integrity, and can be used in a number of downstream applications.
* average yields will vary depending upon a number of factors including species, growth conditions used and developmental stage.
All solutions should be kept tightly sealed and stored at room temperature, except for the RNAse which should be stored at -20 °C.
This kit is designed for research purposes only. It is not intended for human or diagnostic use. Ensure that a suitable lab coat, disposable gloves and protective goggles are worn when working with chemicals. For more information, please consult the appropriate Material Safety Data Sheets (MSDSs). These are available as convenient PDF files online at www.norgenbiotek.com.
Wash Solution I contains guanidinium salts and should be handled with care. Guanidinium salts forms highly reactive compounds when combined with bleach, thus care must be taken to properly dispose of any of these solutions.
You must have the following in order to use the Plant/Fungi DNA Isolation Kit:
All centrifugation steps are carried out in a benchtop microcentrifuge. Various speeds are required for different steps, so please check your microcentrifuge specifications to ensure that it is capable of the proper speeds. All centrifugation steps are performed at room temperature. The correct rpm can be calculated using the formula:
where RCF = required gravitational acceleration (relative centrifugal force in units of g); r = radius of the rotor in cm; and RPM = the number of revolutions per minute required to achieve the necessary g-force.