ABSTRACT
In more recent times, techniques such as polymerase chain reaction (PCR) and Sanger sequencing have allowed scientists to better comprehend the phylogenetic and functional characteristics of microorganisms. Techniques that operate by extracting nucleic acids rely on enzymatic amplification of certain genes from the complex genomic DNA of environmental samples. This chapter examines quantitative real-time PCR techniques that are available in order to identify the advantages and disadvantages due to the use of the most common molecular biology techniques that are currently applied to examine environmental samples for research purposes. Unlike traditional PCR, which is based on end-point detection of amplified genes, q-PCR uses fluorescence-based detection, such as SYBR Green or fluorescent probes, to measure the accumulation of amplicons in real time during each cycle of the PCR. Quantitative PCR (qPCR) is also often used in environmental research for determining gene and transcript numbers that are present within environmental samples.
