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 molecular biology technique used to detect the interaction of a DNA binding protein with its cognate binding sequence. Labeled DNAs were reacted with crude cell extracts and the complexes are run through a non-denaturing polyacrylamide gel. The migration of the labeled DNA through the gel will be slower by being bound. This shift in electrophoretic mobility indicates functional binding between the protein and the DNA.
Diagnostic Methods
Technology
A laboratory test to determine the lowest concentration of a substance that will inhibit a biological or biochemical function, such as microorganism growth or enzymatic activity.
Diagnostic Methods
Technology
A type of spectroscopy that utilizes a laser beam to irradiate a sample containing particles in suspension, resulting in light scattering. Rapid fluctuations in scattering intensity, around a mean value at a certain angle, occur because of particle diffusion and are dependent upon on particle size. The calculated correlation function yields a diffusion coefficient, for a given temperature and viscosity, which can be used to calculate particle size.
Diagnostic Methods
Technology
Analysis that looks for differences in the lengths of microsatellite repeats between alleles. Polymorphic microsatellites are present in the genome every few thousand base pairs and can function as molecular markers.
Diagnostic Methods
Technology
The process of arranging protein, DNA or RNA sequences to identify regions with similar sequences that may elucidate functional, structural, or evolutionary relationships between the sequences.
Diagnostic Methods
Technology
A laboratory test that produces a quantitative measurement of the reflection or transmission properties of a material as a function of wavelength.
Diagnostic Methods
