We use the whole genome shotgun strategy to establish the sequence of a clone larger than the average read length. The idea is to pepper the DNA with sequence reads such that they overlap, and yield, when assembled, the complete sequence of the clone.

Procedures

The clone is prepared, randomly sheared into small pieces and subcloned into a ‘universal’ cloning vector. The library of subfragments is sampled at random, and a number of sequence reads are generated using universal primers directing sequencing from within the cloning vector. These sequence reads are then checked and assembled into contigs by using professional program, and the complete sequence of the clone generated.

Specifications:

Generation of shotgun libraries of 1.6 ~ 3.0 kb DNA pieces with sonication or random enzyme-digestion

Sequencing up to a coverage of 8 ~ 10 fold

Gap closure by various approaches, such as re-sequencing of selected shotgun clones, sequencing of cosmid-ends, primer walking steps, etc.

Expecting a sequence quality of better than 99.99% accuracy

Turn-around time of approximate 3 ~ 4 months after receiving the sample for sequencing the bacterial genome of ~ 30 kb

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.