By Antonio Alonso

This quantity within the tools in Molecular Biology sequence covers tools in forensic DNA profiling, together with protocols for profiling of autosomal STRs, Y-STRs, X-STRs, autosomal SNPs, INDELS, Y-SNPs, mtDNA-SNPs, and mtDNA hypervariable areas HV1 and HV2.

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Primer Set Preparation 1. Fluorescently labeled forward and unlabeled reverse primers are delivered lyophilized and must be reconstituted with TE−4 buffer (see Note 8) to the following concentrations: Unlabeled reverse primers = 200 μM. Labeled forward primers = 100 μM. 2. The unlabeled reverse primers come with specific quantification information listed in pmol and are reconstituted based on this value. Once reconstituted, they are quantified using a UV spectrophotometer absorbance reading at 260 nm.

4. Deoxyribonucleotide Triphosphates A stock solution containing 10 mM of each deoxyribonucleotide triphosphate (dNTP) was purchased from USB Corporation (Cleveland, OH) (see Note 2). 5. Bovine Serum Albumin Bovine serum albumin (BSA) fraction V lyophilized powder, molecular biology grade, nonacetylated was purchased from SigmaAldrich (St. Louis, MO). 2 mg/mL was prepared in nanopure water. 6. Thermal Cycler A GeneAmp® PCR System 9700 (Applied Biosystems) was used to amplify the NC01 miniSTR multiplex.

Goodwin W, Ballard D, Simpson K, Thacker C, Syndercombe Court D, Gow J. Case study: paternity testing – when 21 loci are not enough. In: Doutremepuich C, Morling N, editors. Progress in forensic genetics 10. Amsterdam, The Netherlands. International Congress Series. 2004;1261:460–2. 7. Wenk RE. Incest indices from microsatellite genotypes of mother-child pairs. Transfusion 2008;48:341–8. 8. Hill CR, Kline MC, Coble MD, Butler JM. Characterization of 26 miniSTR loci for improved analysis of degraded DNA samples.

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