Theory & Practices of Clinical Laboratory
Non-Invasive Prenatal Testing (NIPT) has changed the way prenatal care is done. It provides expectant parents with important genetic information about their unborn child through a simple blood test. This innovative screening method analyzes small fragments of DNA from the fetus that are present in the mother’s blood to detect …
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 …
Quantitative real-time PCR (qPCR) has become a new normal in molecular biology for evaluating the gene expression. This technique allows for sensitive and specific detection of mRNA levels, forming the basis of transcriptomic studies. However, because RNA is dynamic and transient, especially when compared to the more stable DNA, optimizing …
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 …