RNA extraction from plant tissues: the use of calcium to precipitate contaminating pectic sugars.
Several protocols and commercial kits are used for the extraction of nucleic acids from different plant tissues. Although there are several procedures available to remove sugars, which hinder the extraction of clean genomic DNA, there are few to assist with extraction of RNA. Those presently used include precipitations with ethylene glycol monobutyl ether or lithium chloride (LiCl), or centrifugation in cesium chloride (CsCl) gradients, but these generally either do not allow high recovery of RNA, are time consuming, rely on hazardous chemicals or need special equipment. Here we present the use of the simple cation, Ca2+, which has been tested and shown to be very efficient for the precipitation of high molecular weight pectic sugars during RNA extraction. Results are presented for different plant tissues, especially tissues of peach and apple fruits at varying ripening stages.
Dal Cin
Valeriano
V
University of Padua, Department of Environmental Agronomy and Crop Science, 35020, Agripolis, Legnaro, Italy. valeriano.dalcin@unipd.it
Danesin
Marcello
M
Rizzini
Fabio Massimo
FM
Ramina
Angelo
A
D002118:Calcium / Q000737:chemistry / Q000494:pharmacology*; D011232:Chemical Precipitation; D005638:Fruit / Q000737:chemistry / Q000235:genetics; D010368:Pectins / Q000737:chemistry* / Q000302:isolation & purification*; D018749:RNA, Plant / Q000235:genetics / Q000302:isolation & purification*; D027824:Rosaceae / Q000737:chemistry / Q000235:genetics*
D018749:RNA, Plant; D010368:Pectins; D002118:Calcium