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Journal of Biological Sciences
  Year: 2009 | Volume: 9 | Issue: 1 | Page No.: 78-82
DOI: 10.3923/jbs.2009.78.82
Effect of Lysis Strategy in Accuracy and Repeatability of Sex Determination by Single Cell Polymerase Chain Reaction Method
F. Farzaneh Dehkordi, A.M. Ahadi, A. Shirazi and M. Sadeghizade

Abstract:
The main objective of this study was to evaluating of different methods applicable to the single cell DNA preparation for embryos sex determination in the early stages. In this study, we have introduced an efficient protocol for DNA extraction step in the single cell PCR method. We compared three different lysis methods including; one physical (nitrogen freezing and thaw) and two chemicals (potassium hydroxide/dithiothreitol; KOH/DTT lysis protocol) and (Nonidet P40/Proteinase K protocol) for utilizing in the single cell PCR procedure. The results of multiplex nested single cell PCR using NP40/PK protocol was eminently more repeatable and efficient for routine applications in the laboratory in compared with the other protocols analyzed in this study. Present results showed that the efficiency of amplification of the target genes in the single blastomer PCR was 94% with NP40/PK protocol, 88% for KOH/DTT lysis protocol and 64% by using of freeze and thaw protocol. We concluded that the NP40/PK protocol was the most efficient method for extracting DNA from a single cell and could be particularly useful for sexing of bovine embryos in the early stages.
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How to cite this article:

F. Farzaneh Dehkordi, A.M. Ahadi, A. Shirazi and M. Sadeghizade, 2009. Effect of Lysis Strategy in Accuracy and Repeatability of Sex Determination by Single Cell Polymerase Chain Reaction Method. Journal of Biological Sciences, 9: 78-82.

DOI: 10.3923/jbs.2009.78.82

URL: http://scialert.net/abstract/?doi=jbs.2009.78.82

 
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