title: An In-House Method for Molecular Monitoring of BCR-ABL.
abstract: OBJECTIVE
  At present, there are a limited number of facilities in Turkey that can provide reliable real-time quantitative(RQ)-PCR BCR-ABL results. The present study aimed to test a cost-effective, in-house method of BCR-ABL quantification,including verification of the method by RQ-PCR validation tests.
  
  
  METHODS
  BCR-ABL and ABL target sequences were cloned into pJET1.2 vectors, from whichcalibrators were prepared and used as templates in RQ-PCR reactions to generate standard curves. Dilutions of K562cells (representing an in vitro simulation of BCR-ABL transcript reduction) were analyzed.
  
  
  RESULTS
  Standard curves were generated from calibrators. These curves were then used to calculate the BCR-ABL andABL copy numbers; in which linear BCR-ABL results were obtained. Repetitive experiments showed that our methodologywas able to detect 1 BCR-ABL positive cell from amnong 1x105 cells.
  
  
  CONCLUSIONS
  The method described herein is suitable for implementation with any RQ-PCR instrument and/or kit forquantify BCR-ABL transcripts.
  
  
  BACKGROUND
  None declared.
authors.affiliation: Dokuz Eylül University, School of Medicine, Department of Medical Biology and Genetics, İzmir, Turkey.
authors.forename: Hakkı Ogün
authors.initials: HO
authors.lastname: Sercan
authors.affiliation: Dokuz Eylül University, School of Medicine, Department of Medical Biology and Genetics, İzmir, Turkey.
authors.forename: Ilgın
authors.initials: I
authors.lastname: Oztürk
authors.affiliation: Dokuz Eylül University, School of Medicine, Department of Medical Biology and Genetics, İzmir, Turkey.
authors.forename: Ceyda
authors.initials: C
authors.lastname: Calışkan
authors.affiliation: Dokuz Eylül University, School of Medicine, Department of Medical Biology and Genetics, İzmir, Turkey.
authors.forename: Melek
authors.initials: M
authors.lastname: Pehlivan
authors.affiliation: Dokuz Eylül University, School of Medicine, Department of Medical Biology and Genetics, İzmir, Turkey.
authors.forename: Zeynep
authors.initials: Z
authors.lastname: Sercan
keywords: BCR-ABL; Chronic myeloid leukemia; Molecular response; Real-time PCR