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      2. 细胞鉴定简介:

        近年来,大量研究表明 STR 基因分型 方法是进行细胞交叉污染和性质鉴定的最有效和准确的方法之一,STR基因分型应用于细胞鉴定已被ATCC等机构强烈推荐。美国的ATCC 细胞库、德国的DSMZ细胞库以及日本的JCRB细胞库等为STR 分型提供了各细胞株的数据供比对。

        STR基因位点由长度为27个碱基对的短串连重复序列组成,这些重复序列广泛存在于人类基因组中,可作为高度多态性标记,被称为细胞的DNA指纹,其可通过PCR(聚合酶链式反应)来检测。STR基因座位上的等位基因可通过扩增区域内重复序列的拷贝数的不同来区分,在毛细管电泳分离之后可通过荧光检测来识别。随后通过一定的计算方法,即可根据所得的STR分型结果与专业的细胞STR数据库比对从而推算出样品所属的细胞系或可能的交叉污染的细胞系名称。


        实验方案:

        样本要求:

        细胞团>10^6个,或一瓶活细胞,或冻存细胞>10^6个;或DNA浓度>50ng/μl,体积>20μl


        有细胞想鉴定?快戳这里……

        参考文献:

        1.  Chatterjee, R. (2007) Cell biology. Cases of mistaken identity. Science 315, 928-31.

        2.   Ruiz Bravo, N. and Gottesman, M. (2007) Notice regarding authentication of cultured cell lines. This can be viewed online at:

        http://grants.nih.gov/grants/guide/notice-files/NOT-OD-08-017.html

        3.   Yoshino, K. et al. (2006) Essential role for gene profiling analysis in the authentication of human cell lines. Human Cell 19, 43–8.

        4.   Szibor, R. et al. (2003) Cell line DNA typing in forensic genetics—the necessity of reliable standards. Forensic Sci. Int. 138, 37–43.

        5.   Dirks, W.G. et al. (2005) Short tandem repeat DNA typing provides an international reference standard for authentication of human cell lines. ALTEX 22, 103–9.

        6.   Masters, J.R. et al. (2001) Short tandem repeat profiling provides an international reference standard for human cell lines. Proc. Natl. Acad. Sci. USA 98, 8012–7.

        7.   (2001) Verify cell line identity with DNA profiling. ATCC Connection: Newsletter of The American Type Culture Collection 21, 1–2.

        8.   Krenke, B. et al. (2002) Validation of a 16-locus fluorescent multiplex system. J. Forensic Sci. 47, 773–85.

        9.   Levinson, G. and Gutman, G.A. (1987) Slipped-strand mispairing: A major mechanism for DNA sequence evolution. Mol. Biol. Evol. 4, 203–21.

        10. Schlotterer, C. and Tautz, D. (1992) Slippage synthesis of simple sequence DNA. Nucleic Acids Res. 20, 211–5.

        11. Smith, J.R. et al. (1995) Approach to genotyping errors caused by nontemplated nucleotide addition by Taq DNA polymerase. Genome Res. 5, 312–7.

        12. Magnuson, V.L. et al. (1996) Substrate nucleotide-determined non-templated addition of adenine by Taq DNA polymerase: Implications for PCR-based genotyping. BioTechniques 21, 700–9.

        13. Walsh, P.S., Fildes, N.J. and Reynolds, R. (1996) Sequence analysis and characterization of stutter products at the tetranucleotide repeat locus vWA. Nucleic Acids Res. 24, 2807–12.

        14. Moller, A., Meyer, E. and Brinkmann, B. (1994) Different types of structural variation in STRs: HumFES/FPS, HumVWA and HumD21S11. Int. J. Leg. Med. 106, 319–23.

        15. Brinkmann, B., Moller A. and Wiegand, P. (1995) Structure of new mutations in 2 STR systems. Int. J. Leg. Med. 107, 201–3.

        16. Griffiths, R. et al. (1998) New reference allelic ladders to improve allelic designation in a multiplex STR system. Int. J. Legal Med. 111, 267–72.

        17. B?r, W. et al. (1997) DNA recommendations: Further report of the DNA Commission of the ISFH regarding the use of short tandem repeat systems. Int. J. Legal Med. 110, 175–6.

        18. Gill, P. et al. (1997) Considerations from the European DNA Profiling Group (EDNAP) concerning STR nomenclature. Forensic Sci. Int. 87, 185–92.

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