Methods in molecular biology (Clifton, N.J.), 2016
Biological samples recovered for forensic investigations are often degraded and/or have low amoun... more Biological samples recovered for forensic investigations are often degraded and/or have low amounts of DNA; in addition, in some instances the samples may be contaminated with chemicals that can act as PCR inhibitors. As a consequence this can make interpretation of the results challenging with the possibility of having partial profiles and false negative results. Because of the impact of DNA analysis on forensic investigations, it is important to monitor the process of DNA profiling, in particular the amplification reaction. In this chapter we describe a method for the in-house generation and use of internal amplification controls (IACs) with DNA profiling kits to monitor the success of the PCR proces. In the example we show the use of the SGM Plus® kit. These controls can also be used to aid the interpretation of the DNA profile.
Proper interpretation of DNA profiles depends on the quality of the DNA samples, the amplificatio... more Proper interpretation of DNA profiles depends on the quality of the DNA samples, the amplification efficiency and the success of post-PCR processing. Chemicals associated with forensic samples can affect the amplification process while random errors occurring during pipetting and electrokinetic injection can cause variability. This can lead to either a reduced signal or lack of DNA profiles. As recommended by the SWGDAM, laboratories carrying out DNA profiling have to adopt standardise and validate procedures, which lead to high levels of quality assurance and control. During amplification, monitoring is restricted to the use of exogenous controls, which are unable to identify issues associated with individual samples. To address this limitation, four PCR Internal Controls (PJC5) i.e. two Internal Amplification Controls (IACs) and two Internal Non-Amplifiable Control (INAC5), to be used with the AmpFtSTR® SGM Plus® kit were developed. The IACs (90 bp and 410 bp) and INACs (80 bp and...
The REvolutionary Approaches and Devices for Nucleic Acid analysis (READNA) project received fund... more The REvolutionary Approaches and Devices for Nucleic Acid analysis (READNA) project received funding from the European Commission for 4 1/2 years. The objectives of the project revolved around technological developments in nucleic acid analysis. The project partners have discovered, created and developed a huge body of insights into nucleic acid analysis, ranging from improvements and implementation of current technologies to the most promising sequencing technologies that constitute a 3(rd) and 4(th) generation of sequencing methods with nanopores and in situ sequencing, respectively.
Methods in molecular biology (Clifton, N.J.), 2016
Biological samples recovered for forensic investigations are often degraded and/or have low amoun... more Biological samples recovered for forensic investigations are often degraded and/or have low amounts of DNA; in addition, in some instances the samples may be contaminated with chemicals that can act as PCR inhibitors. As a consequence this can make interpretation of the results challenging with the possibility of having partial profiles and false negative results. Because of the impact of DNA analysis on forensic investigations, it is important to monitor the process of DNA profiling, in particular the amplification reaction. In this chapter we describe a method for the in-house generation and use of internal amplification controls (IACs) with DNA profiling kits to monitor the success of the PCR proces. In the example we show the use of the SGM Plus® kit. These controls can also be used to aid the interpretation of the DNA profile.
Proper interpretation of DNA profiles depends on the quality of the DNA samples, the amplificatio... more Proper interpretation of DNA profiles depends on the quality of the DNA samples, the amplification efficiency and the success of post-PCR processing. Chemicals associated with forensic samples can affect the amplification process while random errors occurring during pipetting and electrokinetic injection can cause variability. This can lead to either a reduced signal or lack of DNA profiles. As recommended by the SWGDAM, laboratories carrying out DNA profiling have to adopt standardise and validate procedures, which lead to high levels of quality assurance and control. During amplification, monitoring is restricted to the use of exogenous controls, which are unable to identify issues associated with individual samples. To address this limitation, four PCR Internal Controls (PJC5) i.e. two Internal Amplification Controls (IACs) and two Internal Non-Amplifiable Control (INAC5), to be used with the AmpFtSTR® SGM Plus® kit were developed. The IACs (90 bp and 410 bp) and INACs (80 bp and...
The REvolutionary Approaches and Devices for Nucleic Acid analysis (READNA) project received fund... more The REvolutionary Approaches and Devices for Nucleic Acid analysis (READNA) project received funding from the European Commission for 4 1/2 years. The objectives of the project revolved around technological developments in nucleic acid analysis. The project partners have discovered, created and developed a huge body of insights into nucleic acid analysis, ranging from improvements and implementation of current technologies to the most promising sequencing technologies that constitute a 3(rd) and 4(th) generation of sequencing methods with nanopores and in situ sequencing, respectively.
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Papers by Nathalie Zahra