Preparation of genomic DNA from plant tissue.
This unit describes two methods for preparing genomic DNA from plant tissue. In the first method, plant cells are lysed with ionic detergent, treated with protease, and subsequently purified by cesium chloride (CsCl) density gradient centrifugation. The second method is based upon a series of treatments with the nonionic detergent cetyltrimethylammonium bromide (CTAB) to lyse cells and purify nucleic acid. Nucleic acid is recovered from the final CTAB solution by isopropanol or ethanol precipitation. The first method, although somewhat more lengthy, results in highly purified nucleic acid. The second method requires fewer manipulations, results in very high yields (approximately 10-fold higher per gram fresh tissue depending on species and condition of starting material), and produces DNA that is less pure but nonetheless suitable in quality for use in many molecular biology manipulations.
Richards
E
E
Washington University, St. Louis, Missouri, USA.
Reichardt
M
M
Rogers
S
S
D002499:Centrifugation, Density Gradient; D002586:Cesium / Q000737:chemistry; D000077286:Cetrimonium; D002593:Cetrimonium Compounds / Q000737:chemistry; D002712:Chlorides / Q000737:chemistry; D018744:DNA, Plant / Q000737:chemistry / Q000302:isolation & purification*; D003902:Detergents / Q000737:chemistry; D010944:Plants / Q000235:genetics
D002593:Cetrimonium Compounds; D002712:Chlorides; D018744:DNA, Plant; D003902:Detergents; D002586:Cesium; C028019:cesium chloride; D000077286:Cetrimonium