| 单条固定线虫基因组DNA提取及18S rRNA基因PCR扩增 |
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| 引用本文： | 沈锡权,杨官品,廖梅杰.单条固定线虫基因组DNA提取及18S rRNA基因PCR扩增[J].海洋科学,2005,29(5):33-36. |
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| 作者姓名： | 沈锡权 杨官品 廖梅杰 |
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| 作者单位： | 中国海洋大学,海洋生命学院,山东,青岛,266003;中国海洋大学,海洋生命学院,山东,青岛,266003;中国海洋大学,海洋生命学院,山东,青岛,266003 |
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| 基金项目： | 国家自然科学基金资助项目(40176028) |
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| 摘 要： | 根据线虫18S核糖体RM基因PCB扩增效果比较了丙酮、乙醇、乙醇 0．05mol／L FDTA(pH8．0)和5％海水福尔马林4种固定剂，碱裂解和蛋白酶K处理2种单条线虫基因组DNA提取方法的优劣。用乙醇固定的样品最适合制备RR模板DNA，而5％海水福尔马林固定的样品能最完整地保持样品形态。蛋白酶K处理获得的DNA较碱裂解获得的更适合PCR扩增。结果有助于分子生物学方法在海洋线虫分类、多样性和生态学研究中的应用。
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| 关 键 词： | 线虫 固定 基因组DNA 18S核糖体RNA基因 |
| 文章编号： | 1000-3096(2005)05-0033-04 |
| 收稿时间： | 2004/6/11 0:00:00 |
| 修稿时间： | 2004年6月11日 |
Isolation of fixed nematode individual genomic DNA and amplification of18S ribosomal RNA gene fragments |
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| SHEN Xi-quan,YANG Guan-pin,LIAO Mei-jie.Isolation of fixed nematode individual genomic DNA and amplification of18S ribosomal RNA gene fragments[J].Marine Sciences,2005,29(5):33-36. |
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| Authors: | SHEN Xi-quan YANG Guan-pin LIAO Mei-jie |
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| Abstract: | Based on the lengths and yields of PCR products,eight template DNAs were isolated from marine nematode individuals and stored respectively in four fixing solutions(acetone,absolute alcohol,alcohol with0.05mol/L EDTA(pH8.0)and5%formalin in seawater)using two different genomic DNA isolation methods(alkaline lysis and protease K treatment)were compared for their performances in the amplification of18s ribosomal RNA gene fragments.It was found that absolute alcohol is the best fixing solution for preparing PCR template DNA,and5%formalin in seawater is the best for keeping morphological characters of nematode individuals.The genomic DNA isolated with protease K treatment was better for PCR amplification than that isolated with alkaline lysis.Our results would help approach to taxonomical and ecological studies of free-living marine nematode. |
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| Keywords: | nematode fixation genomic DNA 18S ribosomal RNA gene |
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