DNA haplotyping

 

• Service: We determine the haplotype phase among variations in DNA regions of up to 10 kb in size in customer-supplied genomic DNA samples. Larger regions can be haplotyped by having us analyze multiple overlapping regions. We deliver all electropherograms and clone sequence data, as well as final reports summarizing the genotypes of each haplotype for each sample. We also report the alignment of overlapping regions in order to construct larger haplotype relationships.

• Process: We use long-range PCR to create large amplicons of up to 10 kb in size. These large amplicons are purified by gel electrophoresis and cloned into a special plasmid vector suitable for large inserts. After transformation in E. coli, we then plate, grow, and pick colonies, and perform plasmid preparations on 10 clones. The insert region for each clone is fully sequenced using internal primers. The sequences for each clone are aligned to the reference sequence to create a long sequence.¬Ý The 10 long sequences are then analyzed for differences and grouped into two haplotype groups. For each original genomic sample, the consensus sequences of each of the two haplotypes are reported.