狗獾粪便DNA提取方法的初步探讨

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Abstract

The fresh feces of badger (Meles meles) were collected from the dam of Jing-Hang Canal near Yangzhou,and five DNA extraction methods including amylum-subsiding method,preprocess-organicsolvent extraction method,NaCl modified method,GuSCN-SiO₂ method and QIAamp DNA Stool Mini Kit method were used to extract the DNA from them.The results showed that the highest amplification success rate was in the amylum-subsiding method.In this study,fecal sample was lysed rapidly with digested solution,then DNA was extracted using chloroform/phenol after incubation with proteinase K,following by purification to remove numerous potential PCR inhibitors.The mitochondrial DNA control region and 5 microsatellite loci were amplified.showing that the mitochondrial DNA control region sequenced successfully.Our results suggested that DNA extraction based on this method was reliable.

Key words： badger(Meles meles) faecal DNA extraction PCR amplification

Received: 25 December 2008

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	Articles by authors
	LIU Yu-qing
	LV Shen-jin
	YIN Bao-fa
	FAN Hong-mei
	LIU Yan
	WEI Wan-hong

Cite this article:

LIU Yu-qing,LV Shen-jin,YIN Bao-fa, et al. A preliminary study on the DNA extraction method from faeces of badger (Meles meles)[J]. , 2008, 27(6): 468-473.

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http://www.ecolsci.com/EN/ OR http://www.ecolsci.com/EN/Y2008/V27/I6/468

[1] 郑永烈.1987.狗獾.中国动物志[M]. 兽纲,第八卷,食肉目.北京:科学出版社,214-223.

[2] Taberlet P,Waits LP,Luikart G 1999.Noninvasive genetic sampling:Look before you leap[J]. Trends in Ecology and Evolution,14(8),323-327.

[3] 李明,魏辅文,饶刚,等.2001.非损伤性取样法在保护遗传学研究中的应用[J]. 动物学报,47(3):338-342.

[4] Bhagavatula J and Singh L.2006.Genotyping faecal samples of Bengal tiger Panthera tigris tigris for population estimation[J]. A pilot study BMC Genet,7:48.

[5] 魏辅文,饶刚,李明,等.2001.分子粪便学及其应用--可靠性、局限性和展望[J]. 兽类学报,121(02):143-154.

[6] Hoss M M,Kohn M,Paabo S,et al.1992.Excrement analysis by PCR[J]. Nature,359:199.

[7] Reed J Z,Tollit D J,Thompson PM,et al.1997.Molecular scatology:The use of molecular genetic analysis to assign species,sex and individual identity to seal feces[J]. Molecular Ecology,6 (3):225-234.

[8] 万秋红,方盛国,李建国,等.2005.唐家河自然保护区大熊猫的家庭网络:当代基因流评估[J]. 科学通报,50(16):1738-1745.

[9] Zhan X J,Li M,Zhang Z J,et al.2006.Molecular censusing doubles giant panda population estimate in a key nature reserve[J]. Current Biology,16 (12):451-452.

[10] Bellemain E and Taberlet E 2004.Improves noninvasive genotyping method:Application to brown bears (Ursus arctors)[J]. Molecular Ecology Notes,4(3):519-522.

[11] Deagle B E,Tollit D J,Jarman SN,et al.2005.Molecular scatology as a tool to study diet:Analysis of prey DNA in scats from captive Steller sea lions[J]. Molecular Ecology,14 (6):1831-1842.

[12] Hájková P,Zemanová B,Bryja J,et al.2006.Factors affecting success of PCR amplification of microsatellite loci from otter faeces[J]. Molecular Ecology Notes,6(2):559-562.

[13] Nsubuga A M,Robbins M M,Roeder A D,et al.2004.Factors affecting the amount of genomic DNA extracted from ape faeces and the identification of an improved sample storage method[J]. Molecular Ecology,13(7):2089-2094.

[14] 赵健元,李进华,柳杨,等.2005.短尾猴陈旧粪便中DNA的提取[J]. 兽类学报,25(24):410-413.

[15] 程宏毅,鲍毅新,郑荣泉,等.2006.黑麂粪便DNA提取及其PCR检测[J]. 生态科学,25(2):158-161.

[16] 钟华,赖旭龙,魏荣平,等.2003.一种从大熊猫粪便中提取DNA的改进方法[J]. 动物学报,49(5):670-674.

[17] Marmi J,LOpen-Giráldez F,Macdonald DW,et al.2006.Mitochondrial DNA reveals a strong phylogeographic structure in the badger across Eurasia[J]. Molecular Ecology,15(4):1007-1020.

[18] Carpenter P J,Dawson D A,Greig C,et al.2003.Isolation of 39 polymorphic microsatellite loci and the development of a fluorescently labelled marker set for the Eurasian badger (Meles meles)[J]. Molecular Ecology Notes,3(4):610-615.

[19] Frantzen M J,Silk J B,Ferguson J W H et al.1998.Empirical evaluation of preservation methods for faecal DNA[J]. Molecular Ecology,7(10):1423-1428.

[20] 张保卫,魏辅文,李明,等.2004.大熊猫和小熊猫粪便DNA提取的简易方法[J]. 动物学报,50(3):452-458.

[21] Flagstad O,Syvertsen P O,Stenseth N C,et al.2000.Genetic variability in swayne's hartebest,an endagered antelope of Ethiopia[J]. Conservation Biology,14:254-264.

[22] Ernest H B,Penedo M C T,May B P,et al.2000.Molecular tracking of mountain lions in the Yosemite Valley region in California:genetic analysis using microsatellite and faecal DNA[J]. Molecular Ecology,9(4):433-441.

[23] 张德华,周开亚,孙红英.2004.乙醇保存的动物标本基因组DNA提取方法的比较[J]. 生物学杂志,21(6):46-49.

[24] Femando R,Vidya T N,Dangolla A,et al.2003.Reliable noninvasive genotying:Gantasy or reality[J] ? Journal of Herelity,94(2):115-123.

[25] Murphy M A,Waits L P,Kendall KC.2003.The influence of diet on faecal DNA amplification and sex identification in brown bears Ursus arctos[J]. Molecular Ecology,12 (8):2261-2265.