In this interview, News Med is talking to Gabriela Saldanha about DNA Fragmentation and Sanger Sequencing.

What are Sanger sequencing and DNA-fragment analysis? Where are these analyses used?

Sanger sequencing by capillary electrophoresis is the gold standard for DNA sequencing, even with the advent of next-gen sequencing (NGS). The assay and its interpretation are simple and targeted and are best used for DNA fragments that are 75‒1,000 bases long. The current best practice in many labs is to confirm NGS results with Sanger sequencing, and Sanger sequencing is the most cost-efficient method for sequencing single samples.

Fragment analysis separates and analyzes fragments of DNA amplified by PCR. Short tandem repeat (STR) analysis is a subset of fragment analysis and, in the past, has most often been used in forensic and paternity analyses. However, STR analysis can also support non-forensic applications such as cell line authentication, MLPA, microsatellite analysis and mixed-sample deconvolution.

Image Credit: Promega

What instrumentation is required for Sanger sequencing and fragment analysis? Will most researchers have these instruments in their labs, or will they outsource the work?

A single-base resolution capillary electrophoresis system is standard equipment for carrying out these analyses. In the past, many labs would send samples to core labs for analysis because the equipment was often too big or expensive for labs that only needed to…