| Autor: |
Mattocks, Chris, Tarpey, P.S., Whittaker, J.L., Harvey, J.F., Cross, N.C.P. |
| Předmět: |
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| Zdroj: |
Journal of Medical Genetics; Sep2003 Supplement, Vol. 40, pS79-S79, 1/4p |
| Abstrakt: |
The development of capillary technology has dramatically improved the throughput, read length and quality of sequencing now achievable. However, these advances in hardware design have not been matched by suitable improvements to analysis software in terms of sensitivity, specificity or throughput. To address this we have developed prototype software, based on Comparative Sequence Analysis capable of rapidly and accurately analysing sequence data. The minimum input requirement is a known database reference sequence and raw data from two corresponding sequences in the same orientation. Any number of these analyses can be defined and performed simultaneously. Currently a pre-analysis in GeneScan or GeneMapper (Applied Biosystems) is required to define peaks. This information is used to generate an experimental reference sequence comprising peak colours, positions and heights. Using dynamic vertical alignment this is directly compared with equivalent peaks in the defined test traces. Mutations are called and characterised where the normalised peak height in the test significantly falls relative to the experimental reference. Outputs include signal intensity evaluation, quality scores for the sequence and any mutation calls, as well as a graphical representation of the normalised peak heights. We carried out a pilot study comprising 8 repeats of 28 different fragments (average length ∼300 bp) in both orientations, to determine the consistency of normal data. The second phase was a prospective study of 48 cancer cell lines over 48 targets in both orientations carried out in comparison with commercially available software (Mutation Surveyor, SoftGenetics, Inc). The results from these evaluations will be presented and discussed, and the resulting proprietary software presented. [ABSTRACT FROM AUTHOR] |
| Databáze: |
Complementary Index |
| Externí odkaz: |
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