Efficient Isolation of Total RNA from Areca Leaf for High-throughput Sequencing

Key words: 
• Areca catechu L /
• RNA extraction /
• high-throughput sequencing /
• guanidine isothiocyanate-phenol method

Abstract: Total RNA of leaves of Areca nut（ Areca catechu L） were extracted by using universal plant total RNA extraction kit and the guanidine isothiocyanate-phenol method to develop an optimum total RNA extraction protocol for Areca leaves for high-throughput sequencing. A better quality of Areca leaf total RNA was extracted by the guanidine isothiocyanate- phenol than the universal plant total RNA extraction kit. Furthermore,two methods,heat treatment and phenol-chloroform extraction,were tried to inactivate DNAase I after total RNA was digested by this enzyme. It was found that the total leaf RNA was degraded when heat treated at 70 ℃ for 10 min,whereas high-purity and high concentration of total leaf RNA was obtained through phenol-chloroform extraction. The quality of total RNA with OD₂₆₀/ OD₂₈₀= 1. 98,OD₂₆₀/ OD₂₃₀= 2. 31 and concentration more than 979 mg·L^-1,met the standard requirements for high-throughput RNA sequencing.