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 process to measure the proximity of two fluorochromes whereby the excitation energy of one fluorochrome is released and absorbed by the second fluorochrome. It is used to study interactions and structure of macromolecules. (NCI)
Diagnostic Methods
Technology
Hematoxylin-and-eosin, or H&E, is a routine staining procedure of tissue sections. This staining method uses two separate dyes, one staining the nucleus and the other staining the cytoplasm and connective tissue. Hematoxylin is a dark purplish dye that stains the chromatin (nuclear material) within the nucleus. Eosin is an orangish-pink to red dye that stains the cytoplasmic material including connective tissue and collagen.
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
Quantitative PCR: a method to measure very small amounts of RNA transcripts as cDNA targets. There are two strategies currently used, one is called Competitive RTPCR and the other is called Real-Time PCR.
Diagnostic Methods
Technology
A laboratory procedure in which proteins are first separated across a gel, usually polyacrylamide, according to their isoelectric point, and then separated in a perpendicular direction on the basis of their molecular weight.
Diagnostic Methods
