Comparison of five DNA quantification methods

Six commercial preparations of human genomic DNA were quantified using five quantification methods: UV spectrometry, SYBR-Green dye staining, slot blot hybridization with the probe D17Z1, Quantifiler™ Human DNA Quantification kit and RB1 rt-PCR. All methods measured higher DNA concentrations than expected based on the information by the manufacturers. UV spectrometry, SYBR-Green dye staining, slot blot and RB1 rt-PCR gave 39, 27, 11 and 12%, respectively, higher concentrations than expected based on the manufacturers’ information. The DNA preparations were quantified using the Quantifiler™ Human DNA Quantification kit in two experiments. The measured DNA concentrations with Quantifiler were 125 and 160% higher than expected based on the manufacturers’ information. When the Quantifiler™ human DNA standard (Raji cell line) was replaced by the commercial human DNA preparation G147A (Promega) to generate the DNA standard curve in the Quantifiler™ Human DNA Quantification kit, the DNA quantification results of the human DNA preparations were 31% higher than expected based on the manufacturers’ information. The results indicate a calibration problem with the Quantifiler™ human DNA standard for its use with the Quantifiler™ Human DNA Quantification kit. The possible reasons for the problem are discussed and a solution is suggested. The results emphasise the need for standard reference DNA material and standard methods for DNA quantification.     

Analysis of artificially degraded DNA using STRs and SNPs--results of a collaborative European (EDNAP) exercise

Recently, there has been much debate about what kinds of genetic markers should be implemented as new core loci that constitute national DNA databases. The choices lie between conventional STRs, ranging in size from 100 to 450 bp; mini-STRs, with amplicon sizes less than 200 bp; and single nucleotide polymorphisms (SNPs). There is general agreement by the European DNA Profiling Group (EDNAP) and the European Network of Forensic Science Institutes (ENFSI) that the reason to implement new markers is to increase the chance of amplifying highly degraded DNA rather than to increase the discriminating power of the current techniques. A collaborative study between nine European and US laboratories was organised under the auspices of EDNAP. Each laboratory was supplied with a SNP multiplex kit (Foren-SNPs) provided by the Forensic Science Service, two mini-STR kits provided by the National Institute of Standards and Technology (NIST) and a set of degraded DNA stains (blood and saliva). Laboratories tested all three multiplex kits, along with their own existing DNA profiling technique, on the same sets of degraded samples. Results were collated and analysed and, in general, mini-STR systems were shown to be the most effective. Accordingly, the EDNAP and ENFSI working groups have recommended that existing STR loci are reengineered to provide smaller amplicons, and the adoption of three new European core loci has been agreed.     

Estimating the probability of allelic drop-out of STR alleles in forensic genetics

In crime cases with available DNA evidence, the amount of DNA is often sparse due to the setting of the crime. In such cases, allelic drop-out of one or more true alleles in STR typing is possible. We present a statistical model for estimating the per locus and overall probability of allelic drop-out using the results of all STR loci in the case sample as reference. The methodology of logistic regression is appropriate for this analysis, and we demonstrate how to incorporate this in a forensic genetic framework.     

Timespacing competence: multilingual children's linguistic worlds

Drawing on Kramsch's conceptualization of the multilingual subject and the symbolic self, in this paper, we explore how multilingual children re-signify three intertwined myths about the bilingual student, linguistic diversity and language competence, when, in the researcher-generated activity My linguistic world 2014, they are invited to map and talk about their lived experiences as multiple language users seen in the light of place and movement. By demythifying themselves and their linguistic worlds, the children also raise important questions about the notion of linguistic competence. By perceiving competences from a subjective child perspective, we learn how children do what we call timespacing competence. On that basis, we suggest paying attention to how children themselves timespace competence by focusing (more consistently) on the subjective, social, spatial and temporal dimensions of (knowing) language.     

FAILED STATED CRITICS ON PEOPLE＇S CONSTITUTIONAL RIGHTS IGNORANCE IN MORO- MORO,REGISTER 45 FOREST,MESUJI, LAMPUNG

The idea of constitutionalism requires the notion of limiting the power and rights of the people guaranteed by the Constitution. Indonesia Constitution-Undang-Undang Dasar1945 clearly provides protection upon constitutional rights of citizens. Yet, in some places where agrarian conflicts happened, constitutional rights of citizens ignorance still occurs. One of the longest conflicts in Lampung is the conflict in Register 45forest area, in Mesuji district, Lampung. This conflict has been occurring for more than eleven years and resulting in many victims. It is a conflict about control and management of Industrial Plantation Forests （HTI） involving disputes among investors, people, and government.     

Validation of the AmpF?STR SEfiler Plus™ PCR Amplification kit for forensic STR analysis

Validation of the AmpF?STR^® SEfiler Plus™ PCR Amplification kit with 29 and 30 PCR cycles for forensic STR analysis demonstrated that the kit had fewer artefacts than the AmpF?STR^® SGM Plus™ kit (28 PCR cycles). The SEfiler Plus kit was more sensitive and devoid of colour artefacts, but showed more stutters, drop-ins, drop-outs and allelic imbalances.     

HS/GC/ECD分析生物检材样品中的氰化物

目的 建立生物样品中氰化物的衍生化定性定量分析方法。方法 用氯胺T衍生化,HS/GC/ECD分析衍生物CICN。结果 在1ml血中,添加0.2μg氰化钾,回收率为84.6％,RSD为6.39％;在1g肝中添加0.5μg氰化钾,回收率为67.3％,RSD为5.05％;血中检出限为5ng/ml。结论 所建方法能定性定量分析生物样品中的氰化物。