A simple and efficient method for isolation of DNA in high mucilaginous plant tissues.
A protocol is described for rapid DNA isolation from Malvaceae plant species and different tissues of Bixaceae that contain large amounts of polysaccharides, polyphenols, and pigments that interfere with DNA extractions. The method is a modification of Dellaporta et al. The current protocol is simple, and no phenol-chloroform extraction, ethanol, or isopropranol precipitation is required. The method is based in the incubation of soluble DNA with silica, mix in batch during the extraction. The procedure can be completed in 2 h and many samples can be processed at the same time. DNA of excellent quality was recovered and used for polymerase chain reaction (PCR) amplification, restriction enzyme digestion, and Southern blot analysis. The method was used with healthy Bixa orellana and virus-infected Malvaceae plants.
Echevarría-Machado
Ileana
I
Unidad de Bioquímica y Biología Molecular de Plantas, Centro de Investigación Científica de Yucatán, A. C. Calle 43 No. 130, Col. Chuburná de Hidalgo, CP 97200, Mérida, Yucatán, México.
Sánchez-Cach
Lucila A
LA
Hernández-Zepeda
Cecilia
C
Rivera-Madrid
Renata
R
Moreno-Valenzuela
Oscar A
OA
D018744:DNA, Plant / Q000235:genetics / Q000302:isolation & purification*; D004587:Electrophoresis, Agar Gel; D018745:Genome, Plant / Q000235:genetics; D059828:Plant Cells; D010944:Plants / Q000235:genetics*; D016133:Polymerase Chain Reaction; D015183:Restriction Mapping; D013997:Time Factors
D018744:DNA, Plant