Words similar to fbn

Example sentences for: fbn

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Genomic DNA and total RNA were extracted from fibroblast cultures established from skin biopsies as described [ 14 ] . Long-range RT-PCR amplification of FBN1 cDNA fragments and restriction enzyme digestions were performed as previously reported [ 8 ] . Fragments migrating abnormally in agarose gel electrophoresis were excised, purified with the QIAquick Gel Extraction Kit (QIAGEN) and directly sequenced with fluorescent terminators on an ABI Prism 377 DNA sequencer (Perkin-Elmer).
Therefore, we suspected that these normal amplification products were derived from the normal FBN1 alleles, and that the mutant alleles were not amplified by this approach because they carry gross genomic deletions.
Of more than 200 FBN1 mutations reported, only one is a multi-exon deletion [ 7 ] . It involves deletion of exons 60-62 encoding three contiguous cbEGF-like domains [ 23 ] . This deletion co-segregated with affected status in a 3-generation family.
To search for additional large genomic rearrangements, we carried out Southern analyses of BamH I, Sac I and Bgl II digested genomic DNA from 18 fibroblasts strains that were negative for FBN1 mutation screening by other methods, but had shown abnormal fibrillin biosynthetic profiles by pulse-chase analysis.
The blots were hybridized successively with the three overlapping long-range RT-PCR fragments covering the FBN1 coding region, that were generated from a normal cDNA template as described above.