Biotechnology Bulletin ›› 2021, Vol. 37 ›› Issue (2): 71-79.doi: 10.13560/j.cnki.biotech.bull.1985.2020-0540

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Screening of Reference Genes for Quantitative PCR of Skeletal Muscle Fiber Types in Mice

ZHANG Jing1,2(), XIONG Yan1,2(), HUA Yong-lin2, GUO Yu2, XIONG Xian-rong1,2, ZI Xiang-dong1,2, LI Jian1,2()   

  1. 1. Key Laboratory of Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Utilization,Ministry of Education,Chengdu 610041
    2. College of Animal Science and Veterinary Medicine,Southwest Minzu University,Chengdu 610041
  • Received:2020-05-07 Online:2021-02-26 Published:2021-02-26
  • Contact: XIONG Yan,LI Jian E-mail:zjing2513338252@163.com;xiongyan0910@126.com;jianli_1967@163.com

Abstract:

The purpose of this study is to screen the internal reference genes stably expressed in different types of skeletal muscle fibers,so as to provide a reliable basic data for the study of energy and glucose metabolism in skeletal muscle. In this experiment,gastrocnemius muscle,soleus,tibialis anterior muscle and extensor digitorum longus were collected from 6-week-old female mice. The mRNA levels of Gapdh,Actb,Rer1,Hprt1,Ppia,Rpl7,B2m,Sdha and Rpl27 were detected by RT-qPCR,and their expression stabilities were evaluated by Delta CT,geNorm,NormFinder,BestKeeper and RefFinder. The results showed that the top three genes of stability analyzed by Delta CT method were Rpl7 > Actb > Rer1. The top three genes of stability analyzed by NormFinder method were Rpl7>Rer1>Actb. The top three genes of stability analyzed by BestKeeper method were Rpl7> Gapdh> Actb,and the top three genes of stability analyzed by geNorm method were Actb>B2m>Rpl27. Finally,Rpl7 was the most stable internal reference gene after comprehensively analyzed by RefFinder method,while Ppia was the most unstable by these methods. Therefore,Rpl7 can be used as a stable internal reference gene for the detection of skeletal muscle fiber types in mouse.

Key words: mouse, skeletal muscle fiber, internal reference gene, RT-PCR