摘要
Analyzing functional values of RNA using RNA-seq technology is a hot spot of researches. In order to study the medicinal value of Pholiota adipose from the transcriptome level,it is necessary to extract and isolate RNA samples of high purity and high quality P. adipose. In this study,liquid nitrogen grinding Trizol one-step method was used to extract the total RNA of P. adipose. Quality test and statistical comparative analysis were carried out for RNA extract of liquid nitrogen grinding treated and untreated P. adipose. The results showed that the concentration of RNA in the samples treated with liquid nitrogen was much higher than that of the samples without grinding treatment. The OD260/280 of both was about 2,indicating that the purity of RNA was very high. Besides,the ratio of fluorescence intensity of 25 S and 18 S subunit strips of three replicate samples was 1. 8,1. 9,and 1. 9,close to 2,indicating RNA integrity is good. RIN test results of Agilent 2100 were 9. 1,8. 7,and 9. 3,higher than the standard value 6. 8,further proving the integrity. In sum,liquid nitrogen grinding Trizol one-step method is a very simple and efficient method for extracting high quality total RNA of P. adipose.
Analyzing functional values of RNA using RNA-seq technology is a hot spot of researches. In order to study the medicinal value of Pholiota adipose from the transcriptome level,it is necessary to extract and isolate RNA samples of high purity and high quality P. adipose. In this study,liquid nitrogen grinding Trizol one-step method was used to extract the total RNA of P. adipose. Quality test and statistical comparative analysis were carried out for RNA extract of liquid nitrogen grinding treated and untreated P. adipose. The results showed that the concentration of RNA in the samples treated with liquid nitrogen was much higher than that of the samples without grinding treatment. The OD260/280 of both was about 2,indicating that the purity of RNA was very high. Besides,the ratio of fluorescence intensity of 25 S and 18 S subunit strips of three replicate samples was 1. 8,1. 9,and 1. 9,close to 2,indicating RNA integrity is good. RIN test results of Agilent 2100 were 9. 1,8. 7,and 9. 3,higher than the standard value 6. 8,further proving the integrity. In sum,liquid nitrogen grinding Trizol one-step method is a very simple and efficient method for extracting high quality total RNA of P. adipose.
