MUTATION DISCOVERY
The mutation discovery service will screen a sample population for genetic variation in specific areas of the genome and report the genotype of all samples studied at the identified mutation sites. The service can include DNA extraction from blood or tissue for the samples to be screened or use purified genomic DNA as starting material.
Mutation discovery is done by comparative PCR sequencing which consists in amplifying fragments of DNA by PCR (up to about 800bp at a time) followed by bidirectional sequencing. Each read is compared to a published reference sequence for the fragment of interest. If the scope of the project includes regions larger than 800bp, they will be covered by multiple overlapping fragments sequenced separately. To screen 2kb of the promoter of a given gene, three fragments might be required; two small exons separated by a short intron could be covered by a single fragment; or each exon might require its own fragment to be analyzed.
The service includes the designing and ordering of all primers required for the amplification and sequencing of the regions of interest as well as PCR optimizations. Not all chromosomal regions may be screened for mutations using comparative PCR sequencing because of unusual GC content, secondary structure or sequence complexity. Even though such regions may not have been identified until after the optimization steps of the procedure, there are no charges for regions on which sequences cannot be obtained.
After the project is completed, DNA extracted from submitted blood or tissue samples as well as any unused portions of submitted DNA samples will be returned. Alternatively, the facility can archive the samples for future mutation discovery projects or other DNA based assays.
Turnaround time varies from three weeks to several months depending on the size of project.
Pricing for the mutation discovery service: