ABSTRACT
Following the isolation of deoxyribonucleic acid (DNA) from biological samples, the quantitation of amplifiable human DNA is a critical next step in the process of DNA analysis. The Quantifiler® Trio kit provides a simple way to accurately estimate the quantity of human and male DNA with concentrations as low as 5 pg/µL or less. Not only can the Quantifiler® Trio kit determine the quantity of human DNA present, but it can also give an indication of the quality of the sample, which is essential information to have in the decision-making process regarding any downstream testing being done. In this chapter, we describe how to prepare and process quantitation reactions using the Quantifiler® Trio kit. We also provide basic information on how to interpret the results.
Subject(s)
DNA Fingerprinting , DNA , Male , Humans , DNA Fingerprinting/methods , DNA/genetics , DNA/analysis , Microsatellite RepeatsABSTRACT
The GlobalFiler™ PCR Amplification Kit is one of the most sensitive kits that exist today that makes the PCR amplification of human DNA possible. PCR amplification using this specific kit makes millions of copies of 24 specific target sequences in the DNA, called markers or loci. This kit is a 6-dye, short tandem repeat (STR) multiplex assay kit that has a synthetic mix of primers and single-stranded oligonucleotides that are combined with DNA samples and then subjected to 29 or 30 cycles of denaturing, annealing, and extension, as per laboratory protocol. Methods for instrument operation will vary depending on the thermal cycler instrument model that is used. Nevertheless, the GlobalFiler™ PCR Amplification Kit has proven to be a very useful tool to DNA analysts, amplifying extremely low quantities of DNA, making it possible to detect partial, if not full, genetic profiles from a wide range of sample types. This chapter discusses the typical preparation and PCR amplification of human forensic DNA samples, using the GlobalFiler™ PCR Amplification Kit.
