Rebecca L. Margraf

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BACKGROUND Port-wine stains (PWS) are capillary malformations, typically located in the dermis of the head and neck, affecting 0.3% of the population. Current theories suggest that port-wine stains are caused by somatic mutations that disrupt vascular development. OBJECTIVES Understanding PWS genetic determinants could provide insight into new treatments.(More)
The 144-kDa ␭2 protein is a structural component of mammalian reovirus particles and contains the guanylyltransferase activity involved in adding 5؅ caps to reovirus mRNAs. After incubation of reovirus T3D core particles at 52°C, the ␭2 protein became sensitive to partial protease degradation. Sequential treatments with heat and chymotrypsin caused(More)
Hybridization probe melting analysis can be complicated by the presence of sequence variation (benign polymorphisms or other mutations) near the targeted mutation. We investigated the use of "masking" probes to differentiate alleles with similar probe melting temperatures. Selected sequence variation was masked by incorporating mismatches (deletion,(More)
The genotype of the infecting hepatitis C virus (HCV) helps determine the patient's prognosis and the duration of treatment. Heteroduplex mobility analysis (HMA) is a rapid, inexpensive method for genotyping of HCV that does not require sequencing. We developed an HMA that uses temperature gradient capillary electrophoresis (TGCE) to differentiate HCV(More)
Although reported gene variants in the RET oncogene have been directly associated with multiple endocrine neoplasia type 2 and hereditary medullary thyroid carcinoma, other mutations are classified as variants of uncertain significance (VUS) until the associated clinical phenotype is made clear. Currently, some 46 non-synonymous VUS entries exist in curated(More)
BACKGROUND The hepatitis C virus (HCV) genotype determines patient prognosis and duration of treatment, but sequencing of the gene is lengthy and labor-intensive. We used a commercially available nucleic acid extraction system to develop a single-tube extraction-to-sequencing (STETS) method for HCV genotyping. METHODS HCV RNA was purified and amplified in(More)
Next-generation sequencing (NGS) for clinical research and diagnostics is expanding as technical complexity and costs decline. It is now possible to diagnose inherited disorders based on whole-genome or exome sequencing of affected and unaffected relatives or even single affected individuals. To facilitate the use of NGS as a diagnostic tool for identifying(More)
Variant discovery for rare genetic diseases using Illumina genome or exome sequencing involves screening of up to millions of variants to find only the one or few causative variant(s). Sequencing or alignment errors create "false positive" variants, which are often retained in the variant screening process. Methods to remove false positive variants often(More)
Multisample, nonindexed pooling combined with next-generation sequencing (NGS) was used to discover RET proto-oncogene sequence variation within a cohort known to be unaffected by multiple endocrine neoplasia type 2 (MEN2). DNA samples (113 Caucasians, 23 persons of other ethnicities) were amplified for RET intron 9 to intron 16 and then divided into 5(More)
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