Theory & Practices of Clinical Laboratory
Separating nucleic acid is an exceedingly essential tool for molecular biology. Until modern technology could be used, nucleic acid separation was a time-consuming and work-intensive process based on multiple extraction and centrifugation steps, often constrained by small yields and low purity of separation products, and not suitable for automation and …
Proper data analysis is crucial to obtaining valid and relevant results from an experimental system but is especially critical when assessing variations in mRNA expression of genes, the so-called transcriptome. Unlike DNA that is present in each cell throughout the life of the organism, RNA is transiently expressed and levels …
The MIQE (minimum information for the publication of quantitative real-time PCR) guidelines reflect a significant milestone for the transition of a robust “gold standard” in the process of turning the accurate quantitative polymerase chain reaction (qPCR) from a testing technique into real-time quantitative polymerase reaction (QPCR). It has allowed legacy …
Basics of PCR During a number of amplification cycles, real-time PCR collects fluorescent signals from reactions in the polymerase chain. Quantitative real-time PCR involves converting the fluorescent signals from reactions into a numerical value for each sample. PCR is normally based on a quantitative relationship between the quantities of target …
The development of methods of DNA sequencing has contributed to the study of organisms’ genomic sequences. Maxam and Gilbert found the binding site for lactose-repressors by copying and sequencing 24 bases. Sanger and Maxam-Gilbert procedures were more widespread previously and are still used to determine the order of the nucleotide …