Construction of full-length cDNA libraries

Shotgun Library Construction:

Shotgun ( (either restricted or physically sheared ) sequencing strategies can be used to sequence a variety of samples including cosmids, BACs, and PACs. MDS begins by producing randomly generated plasmid clones. The random fragments (typically, 1-3 kb in length) are cloned into a standard vector with common promoter/primer sites. Sequence data is obtained from each end of the random clones and assembled to provide up to 100% coverage of the entire clone. MDS's shotgun sequencing includes:

• Starting material - purified mRNA or total RNA
• In-coming Quality Control
• Sequencing strategy and preparation of the DNA to be sequenced
• Production of a randomly generated plasmid library
• Fluorescent sequencing methodologies
• Contig alignment of each gel read
• Hard copy report
• Original electropherograms
• Consensus sequence on diskette
• Strategies for gap filling
• Return of project materials received and produced as part of project (upon request)
• On-going consultation during the project