Isolation of high-quality RNA from apple (Malus domestica) fruit.
It is difficult to isolate sufficient quantities of high-quality RNA from apple fruit. An abundance of polyphenolic compounds and polysaccharides and a relatively low concentration of RNA in the fruit tissue create conditions that hamper RNA isolation when standard techniques are used. We have developed two RNA isolation methods that include an initial homogenization and extraction with acetone or ethanol. These in turn remove the interfering compounds and precipitate the protein and nucleic acids for subsequent RNA extraction. The quality of RNA was satisfactory with both acetone and ethanol preparations; however, the acetone powder produced consistently higher quantities of RNA.
Asif
Mehar
M
Department of Natural Resource Sciences and Landscape Architecture, University of Maryland, College Park, Maryland 20742, USA.
Trivedi
Prabodh
P
Solomos
Theophanes
T
Tucker
Mark
M
D000096:Acetone; D015152:Blotting, Northern; D000431:Ethanol; D005419:Flavonoids / Q000032:analysis; D005638:Fruit / Q000235:genetics*; D027845:Malus / Q000235:genetics*; D010636:Phenols / Q000032:analysis; D059808:Polyphenols; D011134:Polysaccharides / Q000032:analysis; D018749:RNA, Plant / Q000302:isolation & purification*; D020133:Reverse Transcriptase Polymerase Chain Reaction
D005419:Flavonoids; D010636:Phenols; D059808:Polyphenols; D011134:Polysaccharides; D018749:RNA, Plant; D000096:Acetone; D000431:Ethanol