A rapid and efficient method for purifying high quality total RNA from peaches (Prunus persica) for functional genomics analyses

被引:197
作者
Meisel, L
Fonseca, B
González, S
Baeza-Yates, R
Cambiazo, V
Campos, R
Gonzalez, M
Orellana, A
Retamales, J
Silva, H [1 ]
机构
[1] Univ Andres Bello, Fac Ecol & Recursos Nat, Ctr Biotecnol Vegetal, Santiago 217, Chile
[2] Univ Chile, Fac Ciencias, Dept Biol, Lab Genet Mol Vegetal, Santiago, Chile
[3] Univ Chile, Fac Ciencias Fis & Matemat, Santiago, Chile
[4] Univ Chile, Inst Nutr & Tecnol Alimentos, Santiago 11, Chile
[5] INIA Platina, Santiago, Chile
[6] Univ Chile, Fac Ciencias Agronom, Santiago, Chile
关键词
cDNA library; fruit; fruit trees; functional genomics; peach; polysaccharides; Prunus persica; RNA isolation; Rosaceae; RT-PCR;
D O I
10.4067/S0716-97602005000100010
中图分类号
Q [生物科学];
学科分类号
07 ; 0710 ; 09 ;
摘要
Prunus persica has been proposed as a genomic model for deciduous trees and the Rosaceae family. Optimized protocols for RNA isolation are necessary to further advance studies in this model species such that functional genomics analyses may be performed. Here we present an optimized protocol to rapidly and efficiently purify high quality total RNA from peach fruits (Prunus persica). Isolating high-quality RNA from fruit tissue is often difficult due to large quantities of polysaccharides and polyphenolic compounds that accumulate in this tissue and co-purify with the RNA. Here we demonstrate that a modified version of the method used to isolate RNA from pine trees and the woody plant Cinnamomun tenuipilion is ideal for isolating high quality RNA from the fruits of Prunus persica. This RNA may be used for many functional-enomic based experiments such as RT-PCR and the construction of large-insert cDNA libraries.
引用
收藏
页码:83 / 88
页数:6
相关论文
共 20 条
[1]  
Arumuganathan K, 1991, PLANT MOL BIOL REP, V9, P208, DOI [DOI 10.1007/BF02672069, 10.1007/BF02672069]
[2]   A simple procedure for the isolation of high quality RNA from ripening banana fruit [J].
Asif, MH ;
Dhawan, P ;
Nath, P .
PLANT MOLECULAR BIOLOGY REPORTER, 2000, 18 (02) :109-115
[3]  
Ausubel F.A., 1997, CURRENT PROTOCOLS MO, DOI DOI 10.1.4
[4]   ESTIMATING NUCLEAR-DNA CONTENT IN PEACH AND RELATED DIPLOID SPECIES USING LASER FLOW-CYTOMETRY AND DNA HYBRIDIZATION [J].
BAIRD, WV ;
ESTAGER, AS ;
WELLS, JK .
JOURNAL OF THE AMERICAN SOCIETY FOR HORTICULTURAL SCIENCE, 1994, 119 (06) :1312-1316
[5]  
Chang S. J., 1993, Plant Molecular Biology Reporter, V11, P113, DOI 10.1007/BF02670468
[6]  
CHOMCZYNSKI P, 1987, ANAL BIOCHEM, V162, P156, DOI 10.1016/0003-2697(87)90021-2
[7]   Fruit genome projects ripen on the vine [J].
Christensen, D .
NATURE MEDICINE, 2003, 9 (03) :250-250
[8]  
DAS O P, 1990, Methods in Molecular and Cellular Biology, V1, P213
[9]   Genetic linkage map of peach [Prunus persica (L.) Batsch] using morphological and molecular markers [J].
Dirlewanger, E ;
Pronier, V ;
Parvery, C ;
Rothan, C ;
Guye, A ;
Monet, R .
THEORETICAL AND APPLIED GENETICS, 1998, 97 (5-6) :888-895
[10]   Construction of a BAC library and its application to the identification of simple sequence repeats in peach [Prunus persica (L.) Batsch] [J].
Georgi, LL ;
Wang, Y ;
Yvergniaux, D ;
Ormsbee, T ;
Iñigo, M ;
Reighard, G ;
Abbott, AG .
THEORETICAL AND APPLIED GENETICS, 2002, 105 (08) :1151-1158