Rapid STR analysis of single source DNA samples in 2h

Assessment of Four Polymerases for Low Volume, Fast PCR Amplification of STR Loci for DNA Reference Samples

“…Fast PCR and/or direct PCR are fairly recent improvements from the late 2000s/early 2010s that can be utilized by the forensic DNA community to reduce processing time and/or costs for forensic-type and reference samples [1][2][3][4][5][6][7][8][9][10]. Reduced volume PCR amplification has been shown to improve sensitivity and efficiency [11], but the more reduced the total reaction volume becomes, the more intra-locus peak balance decreases, making extremely low volume reactions likely unsuitable for anything other than single-source reference samples.…”

“…Using fast thermal cyclers, fast PCR has been achieved in as little as 19-26 min [1,2,5]. In general, fast PCR profiles have often suffered from low level artifacts, increased n-4 stutter percentage and incomplete adenylation (-A) [1,2,5,[8][9][10].…”

A protocol for direct and rapid multiplex PCR amplification on forensically relevant samples

“…Especially for this latter purpose, the time available for generating DNA information may be limited, for instance in cases of crimes by a serial perpetrator with a high risk of repetition, abduction with danger of life, time-restricted threats against persons or places, or when custody time of suspects is limited for legal reasons. Several approaches can reduce DNA profiling time: (1) the use of a rapid amplification protocol [1][2][3][4]; (2) DNA profiling without prior DNA extraction, directly on a biological stain [5][6][7] or (3) DNA typing at the scene of crime or police station via automated, miniaturized devices [8][9][10]. The first two approaches have the advantage that DNA typing is performed under the strict laboratory conditions befitting forensic casework, and that amplified products are separated at the nucleotide-level on validated genetic analyzers.…”

“…Regarding rapid thermal cycling, excellent progress has been made by shortening various steps in the PCR protocol, using rapid PCR enzymes and fast thermal cyclers [1][2][3][4]. For these experiments pristine DNAs were used and profiles with strong signal intensity, good peak balance between loci and a limited amount of (adenylation) artefacts were obtained.…”

Validation of Low Volume, Fast PCR Amplification of STR Loci for DNA Reference Samples

“…Reducing amplification processing time has been a major focus for forensic DNA testing over the past 10 years, especially through the advent of fast PCR polymerases, direct PCR, and rapid DNA testing [1][2][3][4][5][6][7][8][9][10][11][12]. But such a reduction in processing time has often been subject to an increase in cost due to reagents and/or equipment, and/or prone to a decrease in STR profile quality.…”