J Integr Plant Biol. ›› 2004, Vol. 46 ›› Issue (5): 578-581.

• Research Articles • Previous Articles     Next Articles

Single-Embryo RT-PCR: Application for Gene Expression Dynamics and Some Technical Aspects

ZOU Jian-Wei, SUN Meng-Xiang, YANG Hong-Yuan   

Abstract:

An extended investigation was carried out for further demonstrating single-embryo RT-PCR technique, which was previously established in our laboratory as a useful tool on study of gene expression of Arabidopsis thaliana (L.) Heynh. Based on the previous work some important technical aspects in the application of the technique were mainly discussed in the present report. The dynamic expression of three selected genes, shoot-meristemless (STM), monppteros (MP) and Arabidopsis shaggy-like kinase etha (ASKh), during the embryogenesis were carefully compared. It was revealed that the technique was especially useful for tracing the expression dynamics of specific genes during embryogenesis. However, the quantity of starting material and the duration of DNaseⅠ treatment are critical points for successful application of the technique. The best condition for eliminating the possible influence of genomic DNA on data analysis was also tested and discussed.

Key words: single-embryo RT-PCR, gene expression patterns, embryogenesis, Arabidopsis thaliana

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