Genomic DNA Extraction Method and AFLP Amplification Reaction System of Blaps rhynchopetera (Coleoptera: Tenebrionidae)

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Genomic DNA Extraction Method and AFLP Amplification Reaction System of Blaps rhynchopetera (Coleoptera: Tenebrionidae)

Abstract

The study compared the quality of DNA extracting from muscles, eggs and larvae of Blaps rhynchopetera with CTAB method, improved SDS-protease-K method, SDS-NaCl method and DNA extraction Kit. Some key factors affecting the time of digestion, dilution of pre-amplification product and the amount of selective amplification primer were studied and an optimized AFLP reaction system of B. rhynchopetera was established. High quality genomic DNA could be isolated using four methods from this beetle muscles. DNA extracted with traditional extraction method could keep for longer time than DNA extraction Kit did. The optimal reaction time for enzyme digestion(EcoR Ⅰ/Mse Ⅰ) was 2 hours for each. Products of the pre-amplification diluted 20 fold and 5 ng/35 ng(E+3/M+3) was the best amount of the selective amplification primer.
- Blaps rhynchopetera,
- genomic DNA,
- DNA extraction,
- AFLP

References

[1]	郑先云, 马恩波, 郭亚平. AFLP分子标记技术在昆虫学研究中的应用[J]. 昆虫知识,2003, 40(5): 405-409
[2] Zabeau M, Vos P. Selective restriction fragment amplification: a general method for DNA fingerprinting: European Patent Office,EP0969102 .1993-03-31
[3] 姜玉锁, 刘文忠, 张春香, 等. 中国境内不同地理型东方蜜蜂遗传多样性的AFLP分析[J]. 昆虫学报,2007, 50(2): 144-152
[4] 鲁成, 赵爱春, 向仲怀, 等. 中国野桑蚕遗传多样性的AFLP分析[J]. 动物学研究,2002, 23(2):166-172
[5] Katiyar S, Chandel G, Tan Y, et al. Biodiversity of Asian rice gall midge (Orseolia oryzae Wood Mason) from five countries examined by AFLP analysis[J]. Genome,2000, 43(2): 322-332
[6] 张烈, 钱敏, 代方银,等. 家蚕高密度AFLP连锁图谱的构建[J]. 昆虫学报,2008, 51(3): 246-257
[7] 朱玉芳, 谭远德, 万春玲,等. 家蚕AFLP连锁框架图谱的构建[J]. 昆虫学报,2001, 44(4): 483-493
[8] Bensch S, kesson M. Ten years of AFLP in ecology and evolution: why so few animals[J]. Molecular Ecology,2005, 14(10):2899-2914
[9] Mueller U, Wolfenbarger L. AFLP genotyping and fingerprinting[J]. Trends In Ecology & Evolution,1999, 14(10): 389-394
[10]	[10] Carisio L, Cervella P, Palestrini C, et al. Biogeographical patterns of genetic differentiation in dung beetles of the genus Trypocopris (Coleoptera, Geotrupidae) inferred from mtDNA and AFLP analyses[J]. Journal of Biogeography, 2004, 31(7): 1149-1162
[11]	[11] Ahern R G, Hawthorne D J, Raupp M J. Phylogeography of a specialist insect, Adelges cooleyi: historical and contemporary processes shape the distribution of population genetic variation[J]. Molecular Ecology,2009, 18(2): 343-356
[12]	[12] 杨兆富, 张雅林. AFLP在昆虫系统学研究中的应用[J]. 昆虫分类学报,2005, 27(4): 315-320
[13]	[13] 张民照, 康乐. AFLP标记的特点及其在昆虫学研究中的应用[J]. 昆虫学报,2002, 45(4): 538-543
[14]	[14] 印红, 刘晓丽, 王彦芳, 等. 一种改进的昆虫基因组DNA的提取方法[J]. 河北大学学报:自然科学版,2002, 22(1): 80-82
[15]	[15] 代金霞, 于有志, 郑哲民. 拟步甲昆虫基因组DNA提取的比较研究[J]. 宁夏大学学报:自然科学版,2004, 25(1): 66-68
[16]	[16] 罗氚芸, 刘勇, 李蕾. 云南琵琶甲脂溶性抗菌活性成分的研究[J]. 中草药,1999, 30(8): 566-567
[17]	[17] 林南英, 刘为忠, 李维莉,等. 药用昆虫云南琵琶甲的化学成分研究[J]. 中国民族民间医药杂志,2000, 44(3): 162-164
[18]	[18] 罗氚芸, 刘勇, 李蕾, 等. 云南民间药用琵琶甲无机元素及氨基酸分析[J]. 氨基酸和生物资源,1999, 21(3): 38-40
[19]	[19] 赵敏, 冯颖, 陈晓鸣, 等. 喙尾琵甲(鞘翅目:拟步甲科)形态记述及生物学特性观察[J]. 环境昆虫学报,2009, 31(4): 348-355
[20]	[20] 赵敏, 陈晓鸣, 孙龙, 等. 云南省喙尾琵琶甲分布及生态环境调查[J]. 林业科学研究,2007, 20(3): 356-362
[21]	[21] 冀焕红, 冯颖, 赵敏, 等. 环境因子对喙尾琵甲卵孵化的影响[J]. 林业科学研究,2010, 23(1): 39-44
[22]	[22] 石晶, 谢映平, 薛皎亮, 等. 蚧虫基因组DNA不同提取方法的比较[J]. 昆虫知识,2005, 42(2): 207-211
[23]	[23] Vos P, Hogers R, Bleeker M, et al. AFLP: a new technique for DNA finger printing[J]. Nucleic Acids Research, 1995, 23(21): 4407-4414
[24]	[24] 陈敏, 陶静, 马超德, 等. 沙棘木蠹蛾AFLP引物筛选及反应体系的建立[J]. 北京林业大学学报,2008, 30(4): 116-120
[25]	[25] Bassam B J, Caetano-Anollés G, Gresshoff P M. Fast and sensitive silver staining of DNA in polyacrylamide gels[J]. Analytical Biochemistry, 1991, 196(1): 80-83
[26]	[26] 王兴红, 高宝嘉, 南宫自艳, 等. 松毛虫AFLP反应体系的建立及引物筛选[J]. 河北农业大学学报,2009, 32(2): 84-88
[27]	[27] 郑先云, 宣涛, 龙文敏, 等. 中华稻蝗AFLP反应体系的建立与优化[J]. 动物分类学报,2007, 32(4): 856-860
[28]

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通讯作者: 陈斌, bchen63@163.com

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沈阳化工大学材料科学与工程学院沈阳 110142

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Genomic DNA Extraction Method and AFLP Amplification Reaction System of Blaps rhynchopetera (Coleoptera: Tenebrionidae)

Abstract: The study compared the quality of DNA extracting from muscles, eggs and larvae of Blaps rhynchopetera with CTAB method, improved SDS-protease-K method, SDS-NaCl method and DNA extraction Kit. Some key factors affecting the time of digestion, dilution of pre-amplification product and the amount of selective amplification primer were studied and an optimized AFLP reaction system of B. rhynchopetera was established. High quality genomic DNA could be isolated using four methods from this beetle muscles. DNA extracted with traditional extraction method could keep for longer time than DNA extraction Kit did. The optimal reaction time for enzyme digestion(EcoR Ⅰ/Mse Ⅰ) was 2 hours for each. Products of the pre-amplification diluted 20 fold and 5 ng/35 ng(E+3/M+3) was the best amount of the selective amplification primer.

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Reference (28)

[1]	郑先云, 马恩波, 郭亚平. AFLP分子标记技术在昆虫学研究中的应用[J]. 昆虫知识,2003, 40(5): 405-409
[2]	[2] Zabeau M, Vos P. Selective restriction fragment amplification: a general method for DNA fingerprinting: European Patent Office,EP0969102 .1993-03-31
[3]	[3] 姜玉锁, 刘文忠, 张春香, 等. 中国境内不同地理型东方蜜蜂遗传多样性的AFLP分析[J]. 昆虫学报,2007, 50(2): 144-152
[4]	[4] 鲁成, 赵爱春, 向仲怀, 等. 中国野桑蚕遗传多样性的AFLP分析[J]. 动物学研究,2002, 23(2):166-172
[5]	[5] Katiyar S, Chandel G, Tan Y, et al. Biodiversity of Asian rice gall midge (Orseolia oryzae Wood Mason) from five countries examined by AFLP analysis[J]. Genome,2000, 43(2): 322-332
[6]	[6] 张烈, 钱敏, 代方银,等. 家蚕高密度AFLP连锁图谱的构建[J]. 昆虫学报,2008, 51(3): 246-257
[7]	[7] 朱玉芳, 谭远德, 万春玲,等. 家蚕AFLP连锁框架图谱的构建[J]. 昆虫学报,2001, 44(4): 483-493
[8]	[8] Bensch S, kesson M. Ten years of AFLP in ecology and evolution: why so few animals[J]. Molecular Ecology,2005, 14(10):2899-2914
[9]	[9] Mueller U, Wolfenbarger L. AFLP genotyping and fingerprinting[J]. Trends In Ecology & Evolution,1999, 14(10): 389-394
[10]	[10] Carisio L, Cervella P, Palestrini C, et al. Biogeographical patterns of genetic differentiation in dung beetles of the genus Trypocopris (Coleoptera, Geotrupidae) inferred from mtDNA and AFLP analyses[J]. Journal of Biogeography, 2004, 31(7): 1149-1162
[11]	[11] Ahern R G, Hawthorne D J, Raupp M J. Phylogeography of a specialist insect, Adelges cooleyi: historical and contemporary processes shape the distribution of population genetic variation[J]. Molecular Ecology,2009, 18(2): 343-356
[12]	[12] 杨兆富, 张雅林. AFLP在昆虫系统学研究中的应用[J]. 昆虫分类学报,2005, 27(4): 315-320
[13]	[13] 张民照, 康乐. AFLP标记的特点及其在昆虫学研究中的应用[J]. 昆虫学报,2002, 45(4): 538-543
[14]	[14] 印红, 刘晓丽, 王彦芳, 等. 一种改进的昆虫基因组DNA的提取方法[J]. 河北大学学报:自然科学版,2002, 22(1): 80-82
[15]	[15] 代金霞, 于有志, 郑哲民. 拟步甲昆虫基因组DNA提取的比较研究[J]. 宁夏大学学报:自然科学版,2004, 25(1): 66-68
[16]	[16] 罗氚芸, 刘勇, 李蕾. 云南琵琶甲脂溶性抗菌活性成分的研究[J]. 中草药,1999, 30(8): 566-567
[17]	[17] 林南英, 刘为忠, 李维莉,等. 药用昆虫云南琵琶甲的化学成分研究[J]. 中国民族民间医药杂志,2000, 44(3): 162-164
[18]	[18] 罗氚芸, 刘勇, 李蕾, 等. 云南民间药用琵琶甲无机元素及氨基酸分析[J]. 氨基酸和生物资源,1999, 21(3): 38-40
[19]	[19] 赵敏, 冯颖, 陈晓鸣, 等. 喙尾琵甲(鞘翅目:拟步甲科)形态记述及生物学特性观察[J]. 环境昆虫学报,2009, 31(4): 348-355
[20]	[20] 赵敏, 陈晓鸣, 孙龙, 等. 云南省喙尾琵琶甲分布及生态环境调查[J]. 林业科学研究,2007, 20(3): 356-362
[21]	[21] 冀焕红, 冯颖, 赵敏, 等. 环境因子对喙尾琵甲卵孵化的影响[J]. 林业科学研究,2010, 23(1): 39-44
[22]	[22] 石晶, 谢映平, 薛皎亮, 等. 蚧虫基因组DNA不同提取方法的比较[J]. 昆虫知识,2005, 42(2): 207-211
[23]	[23] Vos P, Hogers R, Bleeker M, et al. AFLP: a new technique for DNA finger printing[J]. Nucleic Acids Research, 1995, 23(21): 4407-4414
[24]	[24] 陈敏, 陶静, 马超德, 等. 沙棘木蠹蛾AFLP引物筛选及反应体系的建立[J]. 北京林业大学学报,2008, 30(4): 116-120
[25]	[25] Bassam B J, Caetano-Anollés G, Gresshoff P M. Fast and sensitive silver staining of DNA in polyacrylamide gels[J]. Analytical Biochemistry, 1991, 196(1): 80-83
[26]	[26] 王兴红, 高宝嘉, 南宫自艳, 等. 松毛虫AFLP反应体系的建立及引物筛选[J]. 河北农业大学学报,2009, 32(2): 84-88
[27]	[27] 郑先云, 宣涛, 龙文敏, 等. 中华稻蝗AFLP反应体系的建立与优化[J]. 动物分类学报,2007, 32(4): 856-860
[28]

Genomic DNA Extraction Method and AFLP Amplification Reaction System of Blaps rhynchopetera (Coleoptera: Tenebrionidae)

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通讯作者: 陈斌, bchen63@163.com

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Genomic DNA Extraction Method and AFLP Amplification Reaction System of Blaps rhynchopetera (Coleoptera: Tenebrionidae)

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