Two-Step RT-qPCR
In two-step RT-qPCR, cDNA synthesis is carried out using random hexamers, oligo-dT primers, and/or gene-specific primers. Reactions using random hexamers and/or oligo-dT primers produce cDNA that is a mixture of various RNA species in the sample. The cDNA synthesis reaction can be scaled up to accommodate higher RNA input, and extraction and precipitation steps can be used to concentrate and/or further purify the cDNA. This allows better control over the amount of cDNA template that is used for qPCR quantitation. Because only a portion of the cDNA product is used in the subsequent qPCR step, remaining cDNA can be stored for future use, or quantitating the expression of multiple genes from a single RNA/cDNA sample.
Two-step RT-qPCR uncouples cDNA synthesis and subsequent qPCR, allowing greater freedom in selecting RT enzymes and qPCR reagents separately. This flexibility can be useful for controlling sequence representation, efficiency, and optimization of difficult RT-qPCR reactions.
Two-step RT-qPCR uncouples cDNA synthesis and subsequent qPCR, allowing greater freedom in selecting RT enzymes and qPCR reagents separately. This flexibility can be useful for controlling sequence representation, efficiency, and optimization of difficult RT-qPCR reactions.
Choose Type:
Application Notes for Two-Step RT-qPCR
Legal Information
Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. All other trademarks are the property of their respective owners. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@neb.com.
This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
Choose Product:
Two-Step RT-qPCR
