4.5 Article

Reference gene selection for real-time RT-PCR normalization in rice field eel (Monopterus albus) during gonad development

Journal

FISH PHYSIOLOGY AND BIOCHEMISTRY
Volume 40, Issue 6, Pages 1721-1730

Publisher

SPRINGER
DOI: 10.1007/s10695-014-9962-3

Keywords

Reference gene; Real-time RT-PCR; Stability; Monopterus albus

Funding

  1. National Department Public Benefit Research Foundation of China [201003076]
  2. Twelfth 5-Year National Key Science and Technology Research Program of China [2012BAD25B06]
  3. National Natural Foundation of China [30970529]
  4. Fundamental Research Funds for the Central Universities of China [2013PY024]

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Real-time reverse transcriptase (RT) polymerase chain reaction (PCR) requires data normalization using an appropriate reference gene in order to obtain more reliable results with biological significance. We cloned a partial sequence of elongation factor-1-alpha (EF1 alpha) and ribosomal protein L17 (RPL17) from Monopterus albus. We investigated the suitability of five commonly used reference genes [18S ribosomal RNA (18S), cytoskeletal protein (beta-actin), glyceraldehyde phosphate dehydrogenase (GAPDH), EF1 alpha and RPL17] as potential quantitative reference genes for normalizing real-time RT-PCR data generated in gonads of different developmental stages and in other tissues of M. albus. Analysis of the data indicated that 18S, beta-actin and GAPDH are not suitable as reference genes because of their levels of variations of expression. EF1 alpha and RPL17 might be suitable as reference genes in the gonads of different developmental stages as well as in other tissues of M. albus.

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