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High-Precision PCR

NEXT GENERATION SEQUENCING

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.

 

Common Problems

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.

How ThermaStop, ThermaGo & ThermaStop-RT

Overcome Common Problems

ThermaStop ThermaGo 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.