title: Effects of RBC removal and TRIzol of peripheral blood samples on RNA stability.
abstract: BACKGROUND
  Purification of mRNA from stored specimens is very important because results from RT-PCR and microarray analyses are largely affected by the quality of mRNA. Moreover, many preanalytical factors during collection, processing, and storage may affect mRNA quality and the expression of peripheral blood mononuclear cells (PBMC). In this study, we evaluate the effects of RBC removal techniques and TRIzol on RNA quality in blood samples.
  
  
  METHODS
  We obtained EDTA-blood samples from 50 adult volunteers, and made 10 pools of buffy coats for comparison between protocols and also evaluated RNA quality of clinical samples in biobank. Use of TRIzol and RBC removal (RBC lysis or cell separation) were evaluated their effect on the quality of mRNA from the stored blood samples.
  
  
  RESULTS
  RNA integrity with TRIzol was significantly better than that without TRIzol (RIN 4.5 vs. 9.2, respectively; P=0.002). The change in RIN of the PBMC separation method was equivalent to that of the RBC lysis method. After 12 months, IL6 mRNA expression from stored clinical samples in cell separation/TRIzol was stable.
  
  
  CONCLUSIONS
  The blood samples frozen in TRIzol after RBC removal preserved RNA quality well. PBMC/TRIzol preservation for storage of blood samples could be a simple protocol for rapid, low-cost biobanking.
authors.affiliation: Department of Laboratory Medicine, Pusan National University Hospital, School of Medicine, Pusan National University, Busan, South Korea.
authors.forename: Jeong-Eun
authors.initials: JE
authors.lastname: Kang
authors.forename: Sang-Hyun
authors.initials: SH
authors.lastname: Hwang
authors.forename: Jin Hyeon
authors.initials: JH
authors.lastname: Lee
authors.forename: Do Youn
authors.initials: DY
authors.lastname: Park
authors.forename: Hyeong-Hoi
authors.initials: HH
authors.lastname: Kim
mesh_terms: D000328:Adult; D002469:Cell Separation; D004912:Erythrocytes / Q000166:cytology*; D006801:Humans; D012313:RNA / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D012313:RNA