, We let the samples for 5 min to allow sample to homogenize at room temperature and they are always kept on ice after that
, We use the Nanodrop1000 but alternative options exist to measure the concentration of RNA like the Qubit3
, This procedure should only be used if you do not do DNAse I column treatment (see Note 8)
, The ratio of skipping could be calculated by quantifying the ratio of integrated density (obtained via ImageJ software) of the skipped band versus skipped + unskipped bands
, 2-mercaptoethanol needs to be manipulated under a chemical hood according to your laboratory regulations
, Hamlet antibody is a mouse monoclonal IgG1 serotype directed against a synthetic peptide: exon 53 (1999-2016) of human dysferlin
, The exon 32 deleted dysferlin band is shorter (26 amino acid less) than the wild type form of dysferlin
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, Induction of exon skipping of the dystrophin transcript in lymphoblastoid cells by transfecting an antisense oligodeoxynucleotide complementary to an exon recognition sequence, Biochem Biophys Res Commun, vol.226, issue.2, pp.445-449, 1996.
, Overview on applications of antisense-mediated exon skipping, Methods Mol Biol, vol.867, pp.79-96, 2012.
, Lariat branch point mutation in the dysferlin gene with mild limb-girdle muscular dystrophy, Neurology, vol.66, issue.7, pp.1114-1116, 2006.
, Efficient bypass of mutations in dysferlin deficient patient cells by antisense-induced exon skipping, Hum Mutat, vol.31, issue.2, pp.136-142, 2010.
URL : https://hal.archives-ouvertes.fr/hal-01610031
, Exon 32 skipping of dysferlin rescues membrane repair in patients' cells, J Neuromuscul Dis, vol.2, issue.3, pp.281-290, 2015.
, Defective membrane repair in dysferlin-deficient muscular dystrophy, Nature, vol.423, issue.6936, pp.168-172, 2003.
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