Simplified Low‐Copy‐Number DNA Analysis by Post‐PCR Purification

  title={Simplified Low‐Copy‐Number DNA Analysis by Post‐PCR Purification},
  author={Pamela J. Smith and Jack Ballantyne},
  journal={Journal of Forensic Sciences},
Frequently, evidentiary items contain an insufficient quantity of DNA to obtain complete or even partial DNA profiles using standard forensic gentotyping techniques. [] Key Method The amplified product was purified using filtration, silica gel membrane, and enzyme mediated hydrolysis purification techniques and evaluated for their effect on fluorescent allelic signal intensity.
Document Title: Development and Evaluation of a Whole Genome Amplification Method for Accurate Multiplex STR Genotyping of Compromised Forensic Casework Samples
The dcDOP-PCR method significantly improved STR allele success when compared to traditional STR analysis (without pre-amplification), producing strong partial or full profiles in many cases where little to no STR data was previously obtained.
dcDegenerate Oligonucleotide Primed-PCR for Multilocus, Genome-wide Analysis From Limited Quantities of DNA
Modifications of the traditional DOP-PCR reaction to include the use of a more degenerate primer, 12 nonspecific cycles, and a proofreading enzyme allows for a more complete, balanced chromosome amplification from limited and/or compromised clinical and biological samples.
Extended PCR conditions to reduce drop-out frequencies in low template STR typing including unequal mixtures.
Utility of amplification enhancers in low copy number DNA analysis
Investigation on low-quantity DNA samples of PCR additives, betaine, DMSO, PEG, and PCRboost®, found that betaine can improve amplification efficiency of LCN DNA samples, specifically by decreasing stutter in a dual locus system.
Maximizing DNA profiling success from sub-optimal quantities of DNA: a staged approach.
Enhanced low-template DNA analysis conditions and investigation of allele dropout patterns.
Reduced reaction volumes and increased Taq DNA polymerase concentration improve STR profiling outcomes from a real-world low template DNA source: telogen hairs
A comparison of four method variations for the amplification of STRs from LTDNA with a widely used, commercially available kit suggests that the LCN method may be preferred over increased PCR cycles for LTDNA analysis, either with or without consensus profiling and statistical modelling.
Higher Capillary Electrophoresis Injection Settings as an Efficient Approach to Increase the Sensitivity of STR Typing
Higher capillary electrophoresis injection settings were used to efficiently improve incomplete STR profiles generated from low‐level DNA samples under standard polymerase chain reaction (PCR) conditions, and an improved profile of the minor component was obtained.


An improved method for post-PCR purification for mtDNA sequence analysis.
When the yield of purified amplicons, quality of the sequence profile, and ease of assay were evaluated, the use of ExoSAP-IT reagent for post-amplification purification was chosen to replace the filtration method.
Reduced volume PCR amplification reactions using the AmpFlSTR Profiler Plus kit.
Reducing the volume of the AmpFlSTR Profiler Plus reagents offered greater sensitivity and improved the chance of obtaining useful results for samples with very low quantities of DNA and multiple source samples and on the downside, amplifications initiated with less than 0.4 ng of DNA exhibited a twofold increase in the standard deviation of peak ratios.
Reliable genotyping of samples with very low DNA quantities using PCR.
An experimental procedure using PCR that provides a reliable genotype at a microsatellite locus using only a few picograms of template DNA is identified and should be systematically used when genotyping nuclear loci of ancient or forensic samples, museum specimens and hair or feces of free ranging animals.
STR DNA typing: increased sensitivity and efficient sample consumption using reduced PCR reaction volumes.
PCR reaction volume reduction can enhance detection and sensitivity while reducing the consumption of irreplaceable crime scene samples, and improve detection limits/sensitivity and lower sample consumption.
STR genotyping and mtDNA sequencing of latent fingerprint on paper.
NIST Mixed Stain Study 3: DNA quantitation accuracy and its influence on short tandem repeat multiplex signal intensity.
There is a causal relationship between DNA measurement accuracy and the efficiency of STR multiplex analysis, and there are large differences in the efficiencies of amplification, separation, and detection among participants using the same nominal measurement systems.