Low Volume STR Amplification Options: Coupling with Standard or Fast PCR, Traditional or Normalized DNA Extraction, and/or Traditional or Alternative Capillary Electrophoresis

STR amplification leads directly to profile development, which is also impacted by DNA extraction and capillary electrophoresis detection. Amplification for forensic human identification purposes is inherently a costly process; reduced volume reactions have long been an effective cost-savings measure. Processing known, high-quality, single-source DNA samples (i.e., buccal samples) allows for the use of even lower reaction volumes. This chapter provides examples of 3 μL and 6 μL reactions for a variety of commercial amplification kits for use with buccal samples, including standard and fast PCR using KAPA2G™ Multiplex Mix. These reactions can be utilized with traditional DNA extracts or those obtained from a normalized extraction with the ChargeSwitch® Forensic DNA Purification Kit. They can be detected via traditional capillary electrophoresis using POP-4™ polymer and a 36 cm array, or an alternative method using POP-6™ polymer and a 22 cm array on the 3130 series Genetic Analyzer instruments. This chapter also includes protocols for the normalized extraction and alternative detection method.