کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
3327860 | 1590640 | 2007 | 7 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Characterization of Aberrant Melting Peaks in Unlabeled Probe Assays
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موضوعات مرتبط
علوم پزشکی و سلامت
پزشکی و دندانپزشکی
انفورماتیک سلامت
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چکیده انگلیسی
An unlabeled probe assay relies on a double-stranded DNA-binding dye to detect and verify target based on amplicon and probe melting. During the development and application of unlabeled probe assays, aberrant melting peaks are sometimes observed that may interfere with assay interpretation. In this report, we investigated the origin of aberrant melting profiles observed in an unlabeled probe assay for exon 10 of the RET gene. It was determined that incomplete 3â² blocking of the unlabeled probe allowed polymerase-mediated probe extension resulting in extension products that generated the aberrant melting profiles. This report further examined the blocking ability of the 3â² modifications C3 spacer, amino-modified C6, phosphate, inverted dT, and single 3â² nucleotide mismatches in unlabeled probe experiments. Although no 3â² blocking modifications in these experiments were 100% effective, the amino-modified C6, inverted dT, and C3 spacer provided the best blocking efficiencies (1% or less unblocked), phosphate was not as effective of a block (up to 2% unblocked), and single nucleotide mismatches should be avoided as a 3â² blocking modification.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: The Journal of Molecular Diagnostics - Volume 9, Issue 3, July 2007, Pages 290-296
Journal: The Journal of Molecular Diagnostics - Volume 9, Issue 3, July 2007, Pages 290-296
نویسندگان
Shale Dames, Rebecca L. Margraf, David C. Pattison, Carl T. Wittwer, Karl V. Voelkerding,