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Citation:

The Method for Liquidambar formosana DNA Extracting and the Optimization of PCR Procedure

  • Received Date: 2003-01-10
  • Three methods(SDS method, CTAB method and high salt low pH method)for DNA extracting were compared. The CTAB method was the best method for Liquidambar formosana DNA extracting. And PCR procedure was optimized about the factors Mg++, DNA density etc. It's proved that the optimum PCR procedure was as follows:pre-denaturating under 94 ℃ for 4 minutes, denaturating under 94 ℃ for 1 minutes, annealing under 36 ℃ for 1 minutes and extending under 72 ℃ for 1.5 minutes. After 40 cycles, the sample was reacted for 8 minutes under 72 ℃. PCR system includes buffer 2 μL,Mg++ 3.0 mmol·L-1, Taq enzyme 1 U, dNTP 2.5 mmol·L-1, primer 0.7 mmol·L-1,DNA 60 ng.
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The Method for Liquidambar formosana DNA Extracting and the Optimization of PCR Procedure

  • 1. Research Institute of Subtropical Forestry, CAF, Fuyang311400, Zhejang, China

Abstract: Three methods(SDS method, CTAB method and high salt low pH method)for DNA extracting were compared. The CTAB method was the best method for Liquidambar formosana DNA extracting. And PCR procedure was optimized about the factors Mg++, DNA density etc. It's proved that the optimum PCR procedure was as follows:pre-denaturating under 94 ℃ for 4 minutes, denaturating under 94 ℃ for 1 minutes, annealing under 36 ℃ for 1 minutes and extending under 72 ℃ for 1.5 minutes. After 40 cycles, the sample was reacted for 8 minutes under 72 ℃. PCR system includes buffer 2 μL,Mg++ 3.0 mmol·L-1, Taq enzyme 1 U, dNTP 2.5 mmol·L-1, primer 0.7 mmol·L-1,DNA 60 ng.

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