Extraction of total RNA from Chrysanthemum containing high levels of phenolic and carbohydrates.
Some standard methods are available in total RNA extraction of plant tissue, which are effective for the ordinary plants. But it is difficult to extract the total RNA from the plants with high levels of phenolic compounds, carbohydrates, or other compounds that bind and/or coprecipitate with RNA. In this paper, a method was described that used the soluble polyvinylpyrrolidone (PVP), ethanol precipitation, phenol extraction and LiCl precipitation. By this method, RNA capable of reverse-transcription and polymerase chain reaction (PCR) amplification was isolated from Chrysanthemum that contains high levels of phenolic compounds and carbohydrates. The OD260/OD280 absorbance ratio is 2, and the RNA is intact. Other specialized RNA extraction methods failed to deliver suitable product. Bands of PCR products are clearly appearing on the polypropylene acyl gel electrophoresis (PAGE) gel, which indicates that the purity, concentration and integrity of total RNA were suitable for the sequent researches.
Daohong
Wang
W
Key Laboratory of Biomechanics and Tissue Engineering under the State Ministry of Education, College of Bioengineering, Chongqing University, Chongqing 400044, PR China.
Bochu
Wang
W
Biao
Li
L
Chuanren
Duan
D
Jin
Zhang
Z
D002241:Carbohydrates / Q000737:chemistry*; D011232:Chemical Precipitation; D031202:Chrysanthemum / Q000737:chemistry*; D004587:Electrophoresis, Agar Gel; D000431:Ethanol / Q000737:chemistry; D008094:Lithium / Q000737:chemistry; D010636:Phenols / Q000737:chemistry*; D010936:Plant Extracts / Q000737:chemistry*; D011205:Povidone / Q000737:chemistry; D012313:RNA / Q000032:analysis / Q000302:isolation & purification*; D020133:Reverse Transcriptase Polymerase Chain Reaction
D002241:Carbohydrates; D010636:Phenols; D010936:Plant Extracts; D000431:Ethanol; D012313:RNA; D008094:Lithium; D011205:Povidone