Abstract
In addition to effective nucleic acid extraction procedures, the accurate determination of concentration is extremely important to the modern molecular diagnosis. Downstream applications often have a specific target or window of nucleic acid concentration that is required in order to perform optimally. While most automated extraction systems ensure that DNA purification is optimized for most protocols, there is variability in systems, specimen types, and microbes. Sometimes, it is essential to know the exact concentration of a nucleic acid sample. Knowing the accurate concentration can help to prevent unnecessary consumption, enhance reproducibility, enhance amplification of difficult targets, and standardize downstream protocols, such as sequencing. Inaccurate quantification can increase variability in downstream assays, which leads to reduced confidence in results. Here, we also discuss commonly used techniques to separate nucleic molecules. Furthermore, the principle of gel electrophoresis as well as the applications of different electrophoresis techniques are discussed in this chapter.
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