qPCR

Real-time polymerase chain reaction (Real-Time PCR), also known as quantitative PCR (qPCR), is a powerful tool for the detection and profiling of gene expression levels. Unlike conventional PCR, the starting materials for Real-Time PCR/qPCR are RNAs, which have to be transformed into DNAs via reverse transcription process.

Compared to conventional PCR, the biggest advantage of Real-Time PCR/qPCR is that it allows real-time monitoring of PCR products at the end of each amplification cycle. Specifically, products of Real-Time PCR/qPCR are labeled with fluorescent dyes (non-specific fluorescent dyes) that can be incorporated into double stranded DNA, or fluorescence-tagged DNA fragments (sequence-specific fluorescent probe) that are complimentary to specific sequences (Figure 1). Therefore, as the amplification process goes on, the accumulation of PCR products can be reflected by the increase of fluorescence intensities. Eventually, the real-time fluorescent signals collected by the qPCR instruments will be available as quantitative data sets.     

Figure 1. Comparison of two commonly used fluorescent reporters for Real-Time PCR/qPCR.  

As a fast, sensitive method with high specificity and flexibility, the Real-Time PCR/qPCR technique has gained great popularity in the plant biology field. Aside from basic studies toward plant gene functions, Real-Time PCR/qPCR has also been employed in multiple industrial purposes, such as detection of genetically modified organisms (GMOs) and testing for phytopathogens. Lifeasible, an industry-leading plant biotechnology company, provides reliable and high quality services for Real-Time PCR/qPCR experiments within the plant system.

With years of experience in plant molecular study, we proudly offer one-stop service for plant gene expression profiling with Real-Time PCR/qPCR (Figure 2). Besides, our plant biology scientists will provide you with strategic and technical support through each step of the assay, including: 

• Isolation of RNA: isolation of RNA and/or mRNA from a variety of plant tissues (leaf, root, stem, flower, etc.) with high quality.
• Reverse transcription: quality-controlled synthesis of cDNA
• Primer design
• Selection of fluorescent reporters
• Selection of house-keeping control genes
• Optimization of expression level calculation formula

Figure 2. The workflow of our one-step service.

We welcome you to contact us for our product and service catalogs, experimental consultation and collaborations. Our customer-centered services guarantee that each project is given the attention it needs, and ensure the maximal output with reduced cost.