Bovine Parentage Genotyping by Highly Multiplexed Next Generation Sequencing

Current cattle industry practices require that registered cattle have complete paternity and parentage determinations which can cost up to $40 per animal. Our proposed MGST method will have the ability to reduce that cost to just the several dollars range. By using the latest methods in massive sequencing provided by current sequencing devices we will be able to achieve this goal. A simple cocktail of small single strand DNA molecules (probes) directs the genotyping which is interrogated by a thermal stable DNA ligase which has excellent discrimination properties. Another common molecular biology method called the polymerase chain reaction then adds sample specific tags and other common sequences to the ligated probes. This creates a complex library of DNA molecules that now contain the samples genotype information. The library is then processed on a massive scale sequencing device which can sequence 40 million molecules. The data is then counted and these counts used to determine the samples genotypes for all the loci being tested. The method will permit up to 100 or more of genotypes per sample and 1000's of samples to be processed in parallel. It is this multiplexing which will dramatically drive down the prices of genotyping operations. While we will only test paternity and parentage in this project the technology can be applied to any DNA bearing organism and will be suitable for directed genotyping in humans, plants, and other commercially relevant organisms.