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Bulletin of Botanical Research ›› 2012, Vol. 32 ›› Issue (3): 320-325.doi: 10.7525/j.issn.1673-5102.2012.03.012

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Total DNA Extraction and RAPD System Optimization of Endangered Plant Populus euphratica and P.pruinosa

JIAO Pei-Pei;WANG Yan-Qin;PANG Xin-An;LI Zhi-Jun;*   

  1. 1.Key Laboratory of Biological Resource Protection and Utilization of Tarim Basin,Xinjiang Production and Construction Group,Alar 843300;2.College of Plant Sciences,Tarim University,Alar 843300
  • Received:1900-01-01 Revised:1900-01-01 Online:2012-05-20 Published:2012-05-20
  • Contact: LI Zhi-Jun
  • Supported by:
     

Abstract: An improved method based on CTAB was used to extract the total DNA from dry leaves of endangered plant Populus euphratica and P.pruinosa, and to research their reaction system of RAPD-PCR. The results showed that the total DNA extracted by the improved CTAB method could be fit for the RAPD analysis of P.euphratica and P.pruinosa. A suitable RAPD-PCR system for P.euphratica and P.pruinosa was developed as follows: 20 μL PCR solution, including 2.5 mmol·L-1 Mg2+, 0.15 mmol·L-1 dNTP, 0.2 μmol·L-1 primer, 100 ng DNA template and 1.5 U Taq DNA polymerase. By testing the stability of the system with 6 polymorphic primer pairs, the results indicated that the developed system could amplify stably and keep a rich polymorphism. These results provided a reference for the further research on both P.euphratica and P.pruinosa at molecular level.

Key words: Populus euphratica, P.pruinosa, DNA extraction, RAPD optimization

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