Technology
A modified polymerase chain reaction (PCR) technique in which a specific primer can amplify its corresponding allele only; this allows for the determination of the presence or absence of a single, specific allele during genotype testing.
Diagnostic Methods
Technology
A type of polymerase chain reaction technique in which the sample is fractionated, within which individual PCR reactions occur in each fraction.
Diagnostic Methods
Technology
A method for amplifying a DNA base sequence using multiple rounds of heat denaturation of the DNA and annealing of oligonucleotide primers complementary to flanking regions in the presence of a heat-stable polymerase. This results in duplication of the targeted DNA region. Newly synthesized DNA strands can subsequently serve as additional templates for the same primer sequences, so that successive rounds of primer annealing, strand elongation, and dissociation produce rapid and highly specific amplification of the desired sequence. PCR also can be used to detect the existence of the defined sequence in a DNA sample.
Diagnostic Methods
Technology
A technique designed to rapidly enrich and amplify targets derived from samples that yield low levels of nucleic acids, such as formalin-fixed paraffin-embedded (FFPE) tissues, for use in next-generation sequencing. Briefly, total nucleic acid or genomic DNA (gDNA) is isolated from fresh or FFPE tissues. Total nucleic acid is converted to double-stranded cDNA (dscDNA) and cleaned by solid-phase reversible immobilization (SPRI). Then the dscDNA or gDNA is subjected to end repair, adenylation and ligation with a universal half-functional adapter. The modified DNA sample is then subjected to two rounds of nested low-cycle PCR, where the first round uses target gene-specific primers and the second round uses nested target gene-specific primers that are 5' tagged with a common sequencing adapter. The resulting target amplicon library is cleaned using SPRI and is suitable for use in next generation genomic assays.
Diagnostic Methods
Technology
A laboratory procedure in which an RNA strand is first transcribed into a DNA complement and then subjected to PCR amplification. Transcribing an RNA strand into a DNA complement is termed reverse transcription and is done by the enzyme reverse transcriptase.
Diagnostic Methods
Technology
A type of nucleic acid amplification test in which sample DNA is exposed to a single temperature in the presence of multiple sets of primers and a polymerase with replication and high strand displacement activity. This test negates the need for thermal cyclers and may be a low cost alternative to PCR.
Diagnostic Methods
Technology
Polymerase chain reaction that uses two sets of primers, in two sequential runs of amplification, with the second set intended to amplify a target within the first run product.
Diagnostic Methods
Technology
An application of PCR that measures the products generated during each cycle of the polymerase chain reaction process in order to determine the starting amount of template in the reaction.
Diagnostic Methods
Technology
An isothermal, in vitro nucleic acid amplification method that allows primer-directed amplification of a target sequence. It utilizes a primer that can be nicked to create a polymerase start site and a DNA polymerase that will displace a downstream strand as it tracks along the template.
Diagnostic Methods
Technology
A technique used to selectively replicate nucleic acid sequences from RNA templates using reverse transcriptase and RNA polymerase. Reverse transcriptase generates cDNA from the region of interest using primers and then RNA polymerase generates copies of the original RNA template.
Diagnostic Methods