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作 者:朱典[1] 裴黎[1] 张颖[1] 徐小玉[1] 杨雪莹[1]
出 处:《刑事技术》2016年第3期254-256,共3页Forensic Science and Technology
基 金:基本科研业务费专项资金发展支撑项目(2015JB001)
摘 要:目的目前涉毒案件中罂粟种属的DNA检验,主要针对的是罂粟叶、籽、根、茎等可获得高纯度DNA的检材,很少涉及罂粟浆液、残根等仅含微量DNA的样本,本文就涉毒案件中罂粟浆液类含微量DNA样本的种属DNA检验方法进行探索。方法使用涉毒案件中的疑似罂粟浆液,分别取悬浮液、离心后上清液以及干燥的沉渣,采用磁珠法提取纯化DNA,使用罂粟种属特异性荧光引物(P14)按照优化的条件进行PCR扩增,并对扩增产物进行检测,对照已知罂粟测序结果确定其种属来源。结果经离心干燥后的沉渣检出了两条清晰的罂粟特异性DNA片段,增加循环次数可提高特异性条带峰值。结论涉毒案件中DNA含量较低的罂粟浆液类检材也可通过DNA检验确定其种属,为日后相关案件中此类检材的种属检验提供参考,具有重要的应用前景。Objective Current genus identificaiton of poppy in drug cases, is achieved mostly through the DNA testing of the plant's parts, such as the leaf, seed, root and stalk, which contain large amounts of DNA. Poppy slurry is not a good sample for such kind of DNA testing as it contains only a small amount of DNA. The aim of this study is to explore the method of poppy genus identification through DNA testing of poppy serous fluid and similar samples.Methods The suspected poppy slurry was disposed to obtain its suspension that was further centrifugated to separate into the supernatant and sediment. The sediment was dried and grinded into powder. From the suspension, supernatant and powdered sediment, DNA was respectively extracted and purified by using the magnetic beads method. PCR amplification was carried out with poppy specific fluorescent primers(P14) under optimized conditions, and the amplification results were detected by gel electrophoresis. Finally, the genus identification was achieved by comparing the results with a known poppy control sequence. Results Two specific DNA fragments were clearly identified from the sediment group, and the peak values of specific bands increased with PCR cycle number ascending.Conclusion The genus of poppy slurry can be identified through its DNA testing, and the result shows a practical value in drug related cases, suggesting a new choice for the genus determination of toxic plant by testing the DNA of its slurry.
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