A Comparison Between Northern Blotting and Quantitative Real-Time PCR as a Means of Detecting the Nutritional Regulation of Genes Expressed in Roots of Arabidopsis thaliana |
| |
Authors: | GAN Yin-bo ZHOU Zhong-jing AN Li-jun BAO Sheng-jie Brian G Forde |
| |
Affiliation: | [1]College of Agriculture and Biotechnology, Zhejiang University, Hangzhou 310029, P.R.China [2]Department of Biological Sciences, Lancaster University, Lancaster LA1 4YQ, UK |
| |
Abstract: | Quantitative real-time PCR (qRT-PCR) has become a routine and robust technique for measuring the expression of genes of interest, validating microarray experiments and monitoring biomarkers. However, concerns have been raised over the accuracy of qRT-PCR in China as well as in the rest of the world. We have previously used qRT-PCR to study the response of ANR1 and other root-expressed MADS-box genes to fluctuations in the supply of nitrate, phosphate and sulphate under hydroponic growth conditions. In this study, we have used both Northern blotting and qRT-PCR analyses to confirm the nutritional regulation of MADS-box genes in Arabidopsis thaliana and test whether both technologies produce the same results. The information obtained indicated that the qRT-PCR results are consistent with those obtained by Northern blotting hybridization for all the tested root-expressed MADS-box genes, in response to different nitrate,phosphate and sulphate growth conditions. Furthermore, our novel results showed that the expressions of AGL12,AGL18, and AGL19 were all down regulated in response to S and P re-supply in both qRT-PCR and Northern blotting analyses. |
| |
Keywords: | Arabidopsis thaliana MADS-box nutrient regulation Northern blotting quantitative real-time PCR |
本文献已被 维普 万方数据 等数据库收录! |
|