Quantitative real-time PCR using the thermal cyclers, such as ABI®Sequence Detection Systems (ABI GeneAmp 5700 and ABI PRISM 7700 and 7900HT)and the LightCycler® system, is based on detection of a fluorescent signal produced proportionally during the amplification of a PCR product. The chemistry is the key to the detection system . A probe (ie, TaqMan) is designed to anneal to the target sequence between the traditional forward and reverse primers. The probe is labeled at the 5'' end with a reporter fluorochrome (usually 6-FAM, HEX or TET) and a quencher fluorochrome (TAMRA) added at any T position or at the 3'' end. The probe is designed to have a higher Tm than the primers, and during the extension phase, the probe must be 100% hybridized for success of the assay. As long as both fluorochromes are on the probe, the quencher molecule stops all fluorescence by the reporter. However, as Taq polymerase extends the primer, the intrinsic 5'' to 3'' nuclease activity of Taq degrades the probe, releasing the reporter fluorochrome. The amount of fluorescence released during the amplification cycle is proportional to the amount of product generated in each cycle.

Advantages

There are many advantages to quantifying DNA or RNA using this technology, foremost being sensitivity and precision. Furthermore turn-around time for data acquisition and analysis by real-time PCR with the 7700 is short. Setup and thermal cycling require less than 3 to 4 hours and data analysis less than 10 additional minutes. Traditional PCR quantification may require several days. Another advantage of providing the quantitative real-time PCR technology in a core facility setting is reduction of labor time and costs. One half-time employee can easily process 96 samples per day, including setup and data analysis. The use of robotics often found in core facilities allows preparation of samples with speed and accuracy.

Advantages

There are many advantages to quantifying DNA or RNA using this technology, foremost being sensitivity and precision. Furthermore turn-around time for data acquisition and analysis by real-time PCR with the 7700 is short. Setup and thermal cycling require less than 3 to 4 hours and data analysis less than 10 additional minutes. Traditional PCR quantification may require several days. Another advantage of providing the quantitative real-time PCR technology in a core facility setting is reduction of labor time and costs. One half-time employee can easily process 96 samples per day, including setup and data analysis. The use of robotics often found in core facilities allows preparation of samples with speed and accuracy.

Advantages

There are many advantages to quantifying DNA or RNA using this technology, foremost being sensitivity and precision. Furthermore turn-around time for data acquisition and analysis by real-time PCR with the 7700 is short. Setup and thermal cycling require less than 3 to 4 hours and data analysis less than 10 additional minutes. Traditional PCR quantification may require several days. Another advantage of providing the quantitative real-time PCR technology in a core facility setting is reduction of labor time and costs. One half-time employee can easily process 96 samples per day, including setup and data analysis. The use of robotics often found in core facilities allows preparation of samples with speed and accuracy.

Applications

The applications for quantitative real-time PCR are innumerable. Detection of genomic or viral DNA in tissues can be a valuable diagnostic tool. Gene expression can be measured after extraction of total RNA and preparation of cDNA by a reverse transcription (RT) step. Setup and analysis are simple and can more easily be extended to the clinical environment than traditional PCR techniques. Another application is an allelic discrimination assay that can detect single-base nucleotide mutations and polymorphisms.

Shanghai Biochip Co., Ltd.（SBC） was founded in Aug. 2001. The existing shareholders of the company include some well-known universities, research institutes, hospitals and enterprises in China. In Feb. 2003, the National Development and Reform Commission authorized SBC to build up and operate the National Engineering Center for biochip at Shanghai project. The company is located in the Zhangjiang High-tech Park Biotech&Pharmaceutical Industrial Base in Pudong Shanghai with a total area of 40,000 square meters, a construction area of over 20,000 square meters, and a current employee number of more than 300 people.

In accordance with company's development strategy and the construction requirements raised by National Development and Reform Commission for the National Engineering Center, SBC is committed to applying and promoting biotechnology development and products for life science research、health-care、 drug discovery and other fields. Through more than 10 years of construction and development, the technology innovation service system is established which is based on the core platforms of the biological sample bank, tissue chips, gene expression chips and the next-generation sequencing; The industrialized system which takes molecular medical diagnosis technology and product development as the core competitiveness has been set up; And three main business areas which include the services for research and drugs R&D, molecular medicine inspection services, and production and sales of molecular diagnostic product.

SBC obeys the value idea of “Scientific innovation, Business orientation, Career development”, follows the construction requirements for the National Engineering Center, and strives to make the company an internationally-advanced national bio-chip industrialization base which integrates the function of technology innovation, achievements transformation, comprehensive service and personal training as a whole. What’s more, the company’s strategic goal is to be an innovative benchmark enterprise among the industry with an level internationally-advanced and has the capability of independent research and development. It would make a contribution to the goal of making Shanghai an influential technology innovation center and also contribute to the domestic field of biomedical science and technology innovation and industrialization in our country.