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To pilot this hypothesis, we performed germline mutation scanning in 48 women (median age 45, 37 85 years) with invasive breast cancers.
Although we did not investigate BRCA gene mutations in all patients in the present study, we performed BRCA1 and BRCA2 gene mutation scanning in 22 patients who had two or more breast cancer patients in their 1st degree relatives.
NGS is now developing as the diagnostic methodology of choice across a range of applications, including mutation scanning in targeted gene panels and WES for congenital disorders, as well as high depth analysis for tumour profiling.
High resolution melting (HRM) is a new methodology for mutation scanning in which the mutation scanning is carried out in the same tube or well in which the sequence is amplified (reviewed in [ 1]).
Mutation scanning in particular is reliant on heteroduplex formation that distorts the shape of the melting curve as compared with a normalised curve of a wild-type reference (Reed et al, 2007).
High resolution melting analysis (HRMA) is a mutation detection method based on the principle that the melting curves of DNA fragments vary depending on base composition, and is a potentially useful technique for fast genotyping and high-throughput mutation scanning in genetic diagnosis [ 11, 12].
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A mutation scan in 10 dogs revealed five single nucleotide polymorphisms in the htr1B coding sequence.
We demonstrate that High Resolution Melting (HRM) analysis can be used in mutation scans in mixed PCR amplicons containing three homoeologous gene fragments.
Cost-effective and rapid methods for screening are highly desirable for mutation scanning, particularly in clinical settings, where eligibility is partly dictated by cost of testing.
Our results strongly suggest that the HRM approach represents a reliable and highly sensitive method for mutation scanning, especially in diagnosis.
High-resolution melting analysis of polymerase chain reaction products for mutation scanning, which began in the early 2000s, is based on monitoring of the fluorescence released during the melting of double-stranded DNA labeled with specifically developed saturation dye, such as LC-Green.
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