Multiplexed cloning

Techniques for the production of individual cDNAs, cloned in bacteria at the rate of one cDNA per bacterium, were multiplexed to produce cDNA libraries , large collections of cDNAs, simply by repeating the cloning procedure with complex cDNA mixtures. These complex cDNA mixtures, now in bacteria, can be surveyed by retrieving and sequencing each individual cDNA, giving a faithful representation of the abundance of each cDNA in the original mixture if sufficient cDNAs are sequenced. 

True paraUehzation of analytical processes has, so far, not been very successful. In such set-ups, only the separation module (in general the column) is set up in parallel, while solvent delivery, injection, detection, and data processing are multiplexed. These systems will no longer be as simple in operation and maintenance as the cloned systems. 

Products/technologies These include proprietary high-throughput multiplex DNA sequencing positional cloning bioinformatics bacterial genomics PathoGenome sequence database. 

The PCR is carried out in tv o reactions vith one FP and RP or as multiplex reaction with all three primers. The size of the generated DNA fragments varies depending on the location of the breakpoint and each neoplastic clone generates a different DNA fragment (usually shorter than 500bp in MBR and more than 600 bp in MCR). 

Clones are auxotrophically selected in top agar and screened for completeness by multiplex PCR. Those that amplify all PCR products are screened for size by restriction digestion and either pulsed-field or field inversion gel electrophoresis (PFGE or FIGE), followed if necessary by Southern blot. Clones can also be sequenced (6), although it is likely to be difficult to isolate significant qnantities of clone DNA separately from yeast chromosomal DNA. 

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