Direct isolation of high-quality low molecular weight RNA of pear peel from the extraction mixture containing nucleic acid.
Low molecular weight RNA (LMW RNA) is generally obtained either from the total RNA or from total nucleic acids solution. Many steps and chemical reagents are involved in traditional methods for LMW RNA isolation where degradation of LMW RNA often occurs, especially for plant materials with high levels of secondary catabolites. In this study, an efficient method was developed to directly isolate pure LMW RNA from pear peel, a material rich in polyphenolics that is covered with a layer of wax. The method was based on polyethylene glycol (PEG) precipitation combining CTAB buffer which is often used to isolate RNA from polysaccharide-rich and polyphenolics-rich materials. The entire procedure could be completed within 6 h and many samples could be processed at the same time. Few and common chemicals are used with this method. Hence, it could be used as an ordinary method in the laboratory. The developed method was further tested by isolating LMW RNA from Arabidopsis. Using the isolated LMW RNA samples, microRNAs were successfully detected and characterized.
Wang
Xin-wei
XW
College of Horticulture, Nanjing Agricultural University, 1st Weigang, Nanjing, 210095, China.
Xiong
Ai-sheng
AS
Yao
Quan-hong
QH
Zhang
Zhen
Z
Qiao
Yu-shan
YS
D000077286:Cetrimonium; D002593:Cetrimonium Compounds / Q000737:chemistry*; D045424:Complex Mixtures / Q000737:chemistry; D005590:Fractional Precipitation / Q000379:methods*; D008970:Molecular Weight; D009696:Nucleic Acids / Q000737:chemistry; D010936:Plant Extracts / Q000737:chemistry* / Q000302:isolation & purification; D011092:Polyethylene Glycols / Q000737:chemistry*; D031989:Pyrus / Q000737:chemistry* / Q000235:genetics*; D012313:RNA / Q000737:chemistry* / Q000302:isolation & purification*
D002593:Cetrimonium Compounds; D045424:Complex Mixtures; D009696:Nucleic Acids; D010936:Plant Extracts; D011092:Polyethylene Glycols; D012313:RNA; D000077286:Cetrimonium