title: The comparison of DNA quantity between full and half volume single cell whole genome amplification by linker-adapter PCR technique.
abstract: BACKGROUND
  Whole genome amplification (WGA) is a very important step in providing sufficient DNA templates for many high-throughput genetic analyzes. WGA approaches can be subdivided into PCR- or non-PCR-based methods. The PCR amplification category includes PEP-PCR, DOP-PCR and linker-adapter PCR, but only the linker-adapter PCR is suitable for application in preimplantation genetic diagnostic screening because it provides the necessary rapid turnaround time.
  
  
  OBJECTIVE
  Evaluate the ability of linker-adapter WGA commercial kits by using half volume compare with full volume of the reagent amplified DNA extracted from single cell fibroblast.
  
  
  METHODS
  Single cell fibroblast was used based on known genetic profiles. The authors reduced the volume of the reagent and compared the DNA yields and fragmented DNA products with yields and products using the original protocol.
  
  
  RESULTS
  Our result did not show a significant difference between the amount of DNA products between full and half volume method (4.72 vs. 4.89 microg, p-value = 0.56). We achieve a slightly different of fragmented length of WGA products, full volume of reagent received slightly longer length than half volume (502.83 vs. 478.30 bp, p-value = 0.19).
  
  
  CONCLUSIONS
  In this study we have shown that the half volume of the reagent of linker-adapter WGA method amplified DNA extracted from single cell fibroblast was comparable DNA yield and DNA fragmented length with the original method. We need further study extrapolate to evaluate the outcome.
authors.affiliation: Reproductive Endocrinology and Infertility Unit, Department of Obstetrics and Gynecology, Faculty of Medicine Ramathibodi Hospital, Mahidol University, Bangkok, Thailand. matchuporn_m@yahoo.com
authors.forename: Matchuporn
authors.initials: M
authors.lastname: Sukprasert
authors.affiliation: Research Center Faculty of Medicine Ramathibodi Hospital, Mahidol University, Bangkok, Thailand.
authors.forename: Sasivimol
authors.initials: S
authors.lastname: Rattanasiri
authors.affiliation: Ronald O. Perelman and Claudia Cohen Center for Reproductive Medicine and Infertility, Weill Cornell Medical College of Cornell University, New York, USA.
authors.forename: Kungpu
authors.initials: K
authors.lastname: Xu
mesh_terms: D053938:DNA Fragmentation; D005347:Fibroblasts / Q000166:cytology; D006801:Humans; D007202:Indicators and Reagents; D009693:Nucleic Acid Hybridization / Q000379:methods; D016133:Polymerase Chain Reaction / Q000379:methods*; D059010:Single-Cell Analysis
chemical_list: D007202:Indicators and Reagents