Successful STR and SNP typing of FTA Card samples with low amounts of DNA after DNA extraction using a Qiagen BioRobot EZ1 Workstation

FTA Cards (GE Healthcare) have been used for more than 4 years in Denmark for the collection of buccal cells as reference samples in crime cases. Semi-automated protocols for STR typing of DNA on punches of FTA Cards are routinely used. In average, full STR profiles were generated from approximately 95% of the FTA Cards with a standard punching protocol, while partial or no STR profile were obtained from 5% of the samples. Here, the Qiagen BioRobot^® EZ1 Workstation (Qiagen) and the EZ1 DNA Investigator Kit (Qiagen) was used to extract DNA from 29 FTA Cards from which a complete STR profile was not generated with the standard punching protocol. All 29 samples were successfully typed with the AmpF?STR^® Identifiler™ PCR Amplification Kit (Applied Biosystems) and with the SNPforID 49plex SNP assay. The lowest amount of DNA that resulted in complete STR and SNP profiles was 80 pg. The STR and SNP profiles were identical to those generated from another sample collected from each of the 29 individuals.     

LC-MS／MS测定尿液中可卡因及其代谢物苯甲酰爱康宁

目的建立尿液中可卡因(cocaine,COC)及其代谢物苯甲酰爱康宁(benzoylecgonine,BZE)的液相色谱-串联质谱分析方法。方法尿液经固相萃取后,用AllurePFP丙基柱分离,以V(甲醇):V(20mmol/L乙酸胺和0.1%甲酸的缓冲溶液)=80∶20为流动相,采用二级质谱多反应监测模式检测COC和BZE。按10mg/kg的剂量对豚鼠腹腔注射可卡因,给药后收集7d尿液。结果尿液中COC和BZE在2.0～100ng/mL质量浓度范围内线性关系良好(r=0.9995),最低检测限(LOD)为0.5ng/mL;回收率大于90%;日内和日间精密度均小于6%;豚鼠尿液中主要检测目标物是BZE,且BZE检测时限也较COC长。结论所建方法灵敏度高,选择性好,适用于尿液中可卡因和苯甲酰爱康宁的检测。     

大白口蘑子实体的化学成分研究

目的 研究大白口蘑(Tricholoma giganteum Massee)子实体的化学成分。方法 大白口蘑子实体的乙酸乙酯部位经过硅胶、RP-18、Sephadex LH-20等多种材料进行分离纯化, 通过现代波谱技术进行结构鉴定。结果 分离鉴定了12个化合物,它们分别为：亚油酸甲酯（1）、亚油酸（2）、麦角甾-4, 6, 8 (14), 22-四烯-3-酮（3）、麦角甾-5, 7, 22-三烯-3β-醇（4）、过氧麦角甾醇（5）、麦角甾-7, 22-二烯-3β, 5α, 9α-三羟基-6酮（6）、麦角甾-7, 22-二烯-3β, 5α-二羟基-6酮（7）、麦角甾-7, 22-二烯-3β, 5α, 6α, 9α-四醇（8）、麦角甾-7, 22-二烯-3β, 5α, 6β, 9α-四醇（9）、麦角甾-7, 22-二烯-3β, 5α, 6β-三醇（10）、3β-O-glucopyranosyl-5α, 6β-dihydroxy-ergosta-7,22-diene（11）、脑苷脂D（12）。结论 化合物1-3、6-12均为首次从该种真菌中分离得到。     

用GC／ECD方法分析海洛因中毒尿液吗啡代谢物

目的　考查尿检材中海洛因的代谢物吗啡和单乙酰吗啡的液液萃取条件、三氟乙酰化和气相色谱电子捕获检测 (GC/ECD)条件。方法　以烯丙吗啡为内标 ,氯仿∶异丙醇 (9∶1)为液相萃取剂萃取尿中的吗啡和单乙酰吗啡 ,采用MBTFA衍生化 (三氟乙酰化 ) ,GC/ECD检测。结果　尿中加样相对回收率吗啡 89% ,单乙酰吗啡 75 % ,最小检测量吗啡 5 0ng ,单乙酰吗啡 10 0ng。通过实验兔的中毒实验 ,对尿检材进行了分析。 结论　所建立的萃取与检测方法分析海洛因中毒尿检材中的吗啡准确、灵敏 ,可用于海洛因的吸毒检验     

Genetic identification of biological samples buried in clay soil during summer and winter seasons

The analysis of a biological samples found in crime scenes can be a challenging task. Minute amount of DNA, degraded DNA and the presence of inhibitors in biological samples, can interfere with the achievement of a complete genetic profile.Chelating resin, silica membranes and silica-coated with paramagnetic resin were the techniques used in this study for the isolation and purification of DNA. Our aim was to find out the best DNA extraction method for application in the presence of biological samples buried in clay soil.     

Green grabbing at the ‘pharm’ gate: rosy periwinkle production in southern Madagascar

The following article contrasts contemporary rosy periwinkle (Catharanthus roseus) extraction in southern Madagascar with original bioprospecting research conducted 50 years ago. My study shows how plant extraction firms have shifted their approaches by creating new labor forms, which devolve risk and increase exploitation in attempts to capture the valuable biogenetic material needed for drug discovery. The periwinkle symbolizes a complex picture of many dynamic barriers to capitalist penetration at work, including the natural, social and political. Over time, these barriers change and act in conjunction to provide a complex commodity chain expressing many exploitative labor relations of green capitalism.     

两种方法提取汗潜手印DNA的比较研究

目的比较M48和DNeasy○R plant Mini两种方法提取汗潜手印DNA的优劣。方法用M48和DNeasy○Rplant Mini两种方法分别提取16对汗潜手印DNA,并进行DNA定量,比较定量结果。结果 M48法明显比plant Mini法提取到的DNA量多（配对t检验：α=0.05,t=3.45,γ=15,0.002     

Proteinase K challenged by a novel protease

Proteinase K is used in forensic DNA extraction methods for cell lysis and degradation of proteins. Here we compare Proteinase K with a novel protease. We conclude that there is no need to exchange Proteinase K in our methods.     

The use of Hemastix® severely reduces DNA recovery using the BioRobot® EZ1

Abstract: The choice of reagents for presumptive tests for blood, and subsequent extraction methodologies, can significantly affect both the quantity and quality of purified DNA. Blood samples directly tested with Hemastix^® yielded <1% of the DNA recovered from untested samples when purified using the Qiagen BioRobot^® EZ1 and EZ1^® DNA Investigator kit. Full short tandem repeat profiles were obtained from both tested and untested samples, suggesting that the Hemastix^® reagent(s) affect DNA binding, rather than produce DNA damage. The Hemastix^® inhibition of DNA yield could be overcome by the addition of MTL buffer to the sample prior to extraction. Laboratories may wish to modify current procedures for extracting blood samples, utilize other extraction/purification methodologies, or inform their submitting agencies to avoid direct exposure of questioned bloodstains to Hemastix^® reagents.