High-Throughput Screening of BAC Libraries Using Illumina Assays
Author Information
Author(s): Luo Ming-Cheng, Xu Kenong, Ma Yaqin, Deal Karin R, Nicolet Charles M, Dvorak Jan
Primary Institution: University of California, Davis
Hypothesis
The impediment in the construction of physical maps of large eukaryotic genomes can be overcome by screening multidimensional pools of BAC clones using the highly parallel Illumina GoldenGate assay.
Conclusion
The Illumina oligonucleotide assay is an efficient tool for screening BAC libraries and anchoring BAC contigs on genetic maps.
Supporting Evidence
- 87.6% of allele-informative oligonucleotide assays successfully clustered BAC pools into positive and negative groups.
- 95% of assays resulted in positive BAC clones being neighbors in single contigs.
- The new pooling strategy allows direct detection of positive BAC clones without manual deconvolution.
Takeaway
This study shows a new way to quickly find specific DNA sequences in large libraries, making it easier to create genetic maps.
Methodology
The study used a five-dimensional pooling strategy and Illumina GoldenGate assays to genotype BAC clones for specific gene sequences.
Limitations
The study's success rate may be affected by the availability of sufficient single-copy SNP loci for assay design.
Digital Object Identifier (DOI)
Want to read the original?
Access the complete publication on the publisher's website