Mixed Sequence Reader: A Program for Analyzing DNA Sequences with Heterozygous Base Calling
Open Access
- 1 January 2012
- journal article
- Published by Hindawi Limited in The Scientific World Journal
- Vol. 2012, 1-10
- https://doi.org/10.1100/2012/365104
Abstract
The direct sequencing of PCR products generates heterozygous base-calling fluorescence chromatograms that are useful for identifying single-nucleotide polymorphisms (SNPs), insertion-deletions (indels), short tandem repeats (STRs), and paralogous genes. Indels and STRs can be easily detected using the currently available Indelligent or ShiftDetector programs, which do not search reference sequences. However, the detection of other genomic variants remains a challenge due to the lack of appropriate tools for heterozygous base-calling fluorescence chromatogram data analysis. In this study, we developed a free web-based program, Mixed Sequence Reader (MSR), which can directly analyze heterozygous base-calling fluorescence chromatogram data in .abi file format using comparisons with reference sequences. The heterozygous sequences are identified as two distinct sequences and aligned with reference sequences. Our results showed that MSR may be used to (i) physically locate indel and STR sequences and determine STR copy number by searching NCBI reference sequences; (ii) predict combinations of microsatellite patterns using the Federal Bureau of Investigation Combined DNA Index System (CODIS); (iii) determine human papilloma virus (HPV) genotypes by searching current viral databases in cases of double infections; (iv) estimate the copy number of paralogous genes, such asβ-defensin 4 (DEFB4) and its paralogHSPDP3.Keywords
Funding Information
- National Science Council (NSC-98-2320-B-182-034-MY3, NSC-100-2221-E-126-011-MY3, DOH99-TD-I-111-TM013, DOH99-TD-C-111-006, NSC-97-3112-B-001-020, CMRPD190571)
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