Detection of MYCN mRNA in neuroblastoma cell lines by quantitative RT-PCR

FENG Chen, TANG Suo-Qin, WANG Jian-Wen, LIU Li-Zhen, GAO Xiao-Ning, LONG Hui

Chinese Journal of Contemporary Pediatrics ›› 2007, Vol. 9 ›› Issue (1) : 47-50.

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PDF(839 KB)
Chinese Journal of Contemporary Pediatrics ›› 2007, Vol. 9 ›› Issue (1) : 47-50.
EXPERIMENTAL RESEARCH

Detection of MYCN mRNA in neuroblastoma cell lines by quantitative RT-PCR

  • FENG Chen, TANG Suo-Qin, WANG Jian-Wen, LIU Li-Zhen, GAO Xiao-Ning, LONG Hui
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Abstract

OBJECTIVE: To examine the feasibility and practicability of quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) with SYBR GREEN I fluorescence for detecting the MYCN mRNA expression in neuroblastoma cell line LA-N-5. METHODS: MYCN mRNA expression in LA-N-5 cells was measured using real time RT-PCR with SYBR GREEN I. Glyceraldehyde phosphate dehydrogenase (GAPDH) was used as internal control. The level of the MYCN mRNA was calculated as MYCN copies/GAPDH copies. RESULTS: Standard curves were linear and showed high correlations (R2>0.99). The ratio of MYCN mRNA copies to GAPDH mRNA copies was calculated based on specific PCR products. The MYCN mRNA level in LA-N-5 cells was obtained (17.4±1.2). CONCLUSIONS: Quantitative RT-PCR with SYBR GREEN I fluorescence may be a sensitive and reliable method for detecting the MYCN mRNA expression. It may also be potential applicable for detecting the MYCN mRNA expression in the small amount neuroblastoma tissues.[Chin J Contemp Pediatr, 2007, 9 (1):47-50]

Key words

Neuroblastoma / MYCN / Quantitative RT-PCR / SYBR GREEN I

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FENG Chen, TANG Suo-Qin, WANG Jian-Wen, LIU Li-Zhen, GAO Xiao-Ning, LONG Hui. Detection of MYCN mRNA in neuroblastoma cell lines by quantitative RT-PCR[J]. Chinese Journal of Contemporary Pediatrics. 2007, 9(1): 47-50
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