Welcome to Bulletin of Botanical Research! Today is Share:

Bulletin of Botanical Research ›› 2009, Vol. 29 ›› Issue (5): 529-533.doi: 10.7525/j.issn.1673-5102.2009.05.004

Previous Articles     Next Articles

Genomic DNA Extraction and AFLP Analysis System Establishment of Caragana microphylla

CHEN Xu-Hui;GAO Yu-Bao*;ZHU Min-Jie;ZHAO Ting-Ting   

  1. (College of Life Science,Nankai University,Tianjin300071)
  • Received:1900-01-01 Revised:1900-01-01 Online:2009-09-20 Published:2009-09-20
  • Contact: GAO Yu-Bao
  • Supported by:

Abstract: The genomic DNA in the fresh leaves of Caragana microphylla was extracted using a modified SDS method, and then an AFLP silver-stained analysis system suitable for C.microphylla was established by optimizing several main factors. We found that the genomic DNA obtained had good purity and integrity for AFLP analysis with mercapto-ethanol in the extracting solution to prevent formation of oxides and PVP to eliminate phenol from genomic DNA. 500 ng genomic DNA was digested for 4 h with EcoRⅠand MseⅠrestriction enzymes, ligated to adaptors for 12 h at 16℃, pre-amplified using EcoRⅠ+1 and MseⅠ+1 primers and then amplified using EcoRⅠ+3 and MseⅠ+3 primers. The amplified products were resolved in denaturing ployacrylamide gels and stained with silver. With this system, clear ploymerphic fingerprints were obtained, which can be taken as a base for future study of AFLP for C.microphylla populations.

Key words: Caragana microphylla, DNA extraction, AFLP, silver-stained

CLC Number: