Overview of Assay
Next Generation sequencing (NGS) is rapidly becoming a dominant force in all areas of biological research and clinical applications. NGS is a term used to describe a variety of high throughput sequencing methods which involves either short-read or long-read sequencing. For short-read sequencing the strategies for preparation of the template varies based on the sequencing system used and the information depth needed. It is a multi-step process that involves PCR or RT-PCR in a variety of different strategies from emulsion PCR, solid-phase bridge amplification and solid-phase template walking to RNA sequencing.
Some of these strategies require PCR steps which uses high fidelity DNA polymerase with complex primer arrays or sequence specific priming which mimics a multiplex PCR situation. While for an RNA target some strategies require gene specific priming (multiplex) and a reverse transcriptase which mimics a “two-step RT-PCR” method. Both are fundamentally the same as any PCR or RT-PCR reaction which can produce non-specific amplification products which reduces the potential yield of the correct product. ThermaStop and ThermaStop-RT are simple-to-use reagents that can reduce or eliminate these unwanted side reactions thus increasing yield of the specific products and increases assay sensitivity.
How ThermaStop and ThermaGo Overcome Common Problems