title: Quantification of DNA Extracted from Formalin Fixed Paraffin-Embeded Tissue Comparison of Three Techniques: Effect on PCR Efficiency.
abstract: BACKGROUND
  Mutation detection from Formalin Fixed Paraffin-Embedding (FFPE) tissue in molecular lab became a necessary tool for defining potential targeted drug. Accurate quantification of DNA extracted from FFPE tissue is necessary for downstream applications like Polymerase Chain Reaction (PCR), sequencing etc.
  
  
  OBJECTIVE
  To check and define which method for FFPE DNA quantification is suitable for downstream processes.
  
  
  METHODS
  In this experimental experience study Biorad Smartspec Plus spectrophotomery, Qubit Fluorometer, and Qiagen Rotorgene qPCR was used to compare 20 FFPE DNA quantification in Rajiv Gandhi Cancer Institute and Research Centre, in 2015 and quantified amount of DNA used for PCR reaction.
  
  
  RESULTS
  The average concentration of DNA extracted from FFPE tissue measured using the spectrophotometer was much higher than the concentration measured using the Qubit Fluorometer and qPCR.
  
  
  CONCLUSIONS
  Results varied depending upon the technique used. A fluorometric analysis may be more suitable for quantification of DNA samples extracted from FFPE tissue compared with spectrophotometric analysis. But qPCR is the best technique because it details DNA quantity along with quality of amplifiable DNA from FFPE tissue.
authors: Ph.D Scholar, Department of Bioengineering & Technology, Gauhati University , Assam, India .
authors: Dushyant
authors: D
authors: Kumar
authors: Ph.D Scholar, Department of Bioengineering & Technology, Gauhati University , Assam, India .
authors: Manoj Kumar
authors: MK
authors: Panigrahi
authors: Head, Centre for Molecular Diagnostic and Cell Biology, Rajiv Gandhi Cancer Institute and Research Centre , New Delhi, India .
authors: Moushumi
authors: M
authors: Suryavanshi
authors: Director Lab Services, Rajiv Gandhi Cancer Institute and Research Centre , New Delhi, India .
authors: Anurag
authors: A
authors: Mehta
authors: Assistant Professor, Department of Bioengineering & Technology, Gauhati University , Assam, India .
authors: Kandarpa Kumar
authors: KK
authors: Saikia
keywords: Fluorometer; Sequencing; Spectrophotometer; qPCR