Selection of housekeeping genes for normalization of RT-PCR in hypoxic neural stem cells of rat in vitro
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Gene expression analysis under various conditions using real-time reverse transcription polymerase chain reaction (RT-PCR) needs reliable control genes. Housekeeping genes are commonly used as the control. However, no validated housekeeping gene is available for study of hypoxic neural stem cell culture. To choose appropriate internal control genes, the expression of eight commonly used housekeeping genes was examined in rat neural stem cell model to find one or more stably expressed genes under hypoxic/ischemic conditions. Two genes, HPRT and RPL13A were identified as the most confidential housekeeping genes in this research by geNorm and NormFinder softwares. As a groundwork, the most stable housekeeping genes for neural stem cells under hypoxic/ischemic conditions are initially investigated and validated in this experiment, which might provide a better understanding for the gene expression study in ischemic and necrotic neural stem cell cultures or in ischemic diseases of the central nervous system (CNS).
KeywordsHousekeeping gene Neural stem cells Hypoxic condition RT-PCR
This work was supported by grants from National Natural Science Foundation of China (30770673 and 81070998) and Youth Fund of Xi’an Jiaotong University College of Medicine (YQN0802).
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