期刊
ANALYTICAL BIOCHEMISTRY
卷 394, 期 2, 页码 202-208出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ab.2009.07.043
关键词
Quantified transcription level; Exogenous RNA and DNA references; Dual-spike-in strategy; Tissue-specific expression; Silkworm Bombyx mori
资金
- State Key Laboratory of Molecular Biology
- National Basic Research Program of China [2005CB121000]
- National Hi-Tech Research and Development Program of China [2006AA10A118]
- National Natural Science Foundation of China [30770497]
To quantify the transcription level of a gene, we have conceived a novel concept, transcription level of messenger RNA (mRNA) per gene copy, which was determined with a dual-spike-in strategy. In this strategy, an exogenous DNA was added as the spike reference for target DNA in addition to the exogenous RNA as the reference for target RNA. After the mRNA-to-DNA ratio of a target gene was estimated by real-time polymerase chain reaction (PCR), it was first normalized with the mRNA-to-DNA ratio of the exogenous reference. The normalized ratio was multiplied by the ratio of exogenous RNA to exogenous DNA to obtain the transcription level of mRNA per gene copy. This quantified transcription value allows one to compare the expression of a target gene in different tissues or the expression in a specified tissue under different conditions. (C) 2009 Elsevier Inc. All rights reserved.
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