title: [Detection of MYCN mRNA in neuroblastoma cell lines by quantitative RT-PCR].
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.
authors.affiliation: Department of Pediatrics, People's Liberation Army General Hospital, Beijing 100853, China.
authors.forename: Chen
authors.initials: C
authors.lastname: Feng
authors.forename: Suo-Qin
authors.initials: SQ
authors.lastname: Tang
authors.forename: Jian-Wen
authors.initials: JW
authors.lastname: Wang
authors.forename: Li-Zhen
authors.initials: LZ
authors.lastname: Liu
authors.forename: Xiao-Ning
authors.initials: XN
authors.lastname: Gao
authors.forename: Hui
authors.initials: H
authors.lastname: Long
mesh_terms: D045744:Cell Line, Tumor; D006801:Humans; D009447:Neuroblastoma / Q000378:metabolism* / Q000473:pathology; D016271:Proto-Oncogene Proteins c-myc / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: D016271:Proto-Oncogene Proteins c-myc; D012333:RNA, Messenger