摘要
The molecular techniques including Northern blot, dot blot, in situ hybridization, etc. have been successfully used to estimate semi-quantitatively mRNA levels in plant samples. In this study, we employed a real-time reverse transcription-PCR (RT-PCR) assay using SYBR Green I fluorescence methodology to evaluate accurate quantitation and sequence specific detection of Aux/IAA mRNA levels in Arabidopsis. Results obtained indicate a linear dynamic range of 102-106 Aux/IAA mRNA copies with standard deviations of generally less than 15%. As a model experiment, the outcome of analysis of expression patterns of five Aux/IAA genes in Arabidopsis under various chemical and temperature treatments is presented. The method presented here provides a sensitive and rapid technique to evaluate plant Aux/IAA mRNA expression levels in nanogram order.
The molecular techniques including Northern blot, dot blot,in situ hybridization, etc. have been successfully used to estimate semi-quantitatively mRNA levels in plant samples. In this study, we employed a real-time reverse transcription-PCR (RT-PCR) assay using SYBR Green I fluorescence methodology to evaluate accurate quantitation and sequence specific detection ofAux/IAA mRNA levels inArabidopsis. Results obtained indicate a linear dynamic range of 102–106 Aux/IAA mRNA copies with standard deviations of generally less than 15%. As a model experiment, the outcome of analysis of expression patterns of fiveAux/IAA genes inArabidopsis under various chemical and temperature treatments is presented. The method presented here provides a sensitive and rapid technique to evaluate plantAux/IAA mRNA expression levels in nanogram order.
基金
J. Liu was supported by the JAERI Research Fellow Program.
