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植物研究 ›› 2006, Vol. 26 ›› Issue (5): 589-594.doi: 10.7525/j.issn.1673-5102.2006.05.016

• 论文 • 上一篇    下一篇

木本植物基因组DNA提取及鉴定

王 军;杨传平*;刘桂丰   

  1. (东北林业大学林学院,哈尔滨 150040)
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2006-09-20 发布日期:2006-09-20
  • 通讯作者: 杨传平
  • 基金资助:
     

Extraction of genomic DNA from woody plants and it’s identification

WANG Jun;YANG Chuan-Ping*;LIU Gui-Feng   

  1. (College of Forestry, Northeast Forestry University, Harbin 150040)
  • Received:1900-01-01 Revised:1900-01-01 Online:2006-09-20 Published:2006-09-20
  • Contact: YANG Chuan-Ping
  • Supported by:
     

摘要: 采用改良后的CTAB法,对山葡萄、软枣猕猴桃、蒙古栎、核桃楸、西伯利亚红松和偃松基因组DNA进行提取。结果表明,所提基因组DNA分子量与λDNA(48 kb)接近,其紫外吸收比在1.66~1.89之间。第3次和第4次上清提取的DNA质量优于第1次和第2次。从提取产量看,每克鲜重提取DNA量最小为15 μg·g-1(核桃楸第4次上清),最高的为272 μg·g-1(山葡萄第3次上清)。西伯利亚红松和偃松第1次和第2次上清基本未提出DNA,第3次和第4次上清中得到了较高质量的DNA。经酶切鉴定和PCR扩增,所提的基因组DNA可以用于进一步研究。

关键词: 木本植物, 基因组DNA, 提取

Abstract: Genomic DNA was extracted from leaves of Vitis amurensis, Actinidia arguta, Quercus mongolica, Juglans mandshurica and needles of Pinus sibirica, Pinus pumila by the method of modified CTAB. The isolated genomic DNA length was similar to λDNA (48 kb) and is suitable for both PCR amplification and digestion with restriction endonucleases. The absorbance ration (A260/A280) ranged 1.66~1.89. The quality of DNA extracted from third and fourth times suspension was better than from first and second times suspension. The yields of DNA ranged from 15 μg·g-1 fresh mass(from fourth time suspension J.mandshuric) to 272 μg·g-1 fresh mass(from third time suspension of V.amurensis).Although from first and second times suspension of P.pumila and P.sibirica wasn’t isolated genomic DNA, but from third and fourth times suspension was obtained high quality genomic DNA. The genomic DNA can be used for next analysis via identification of PCR and digestion with restriction endonucleases.

Key words: woody plants, genomic DNA, extraction

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