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However, this has been excluded by sequencing all samples in three regions (C2Hs, C10Hs and C11Hs).
Ninety-two patinnts in whom the known monogenic causes of obesity had been excluded by sequencing of the genes encoding LEP, LEPR, POMC and MC4R were selected for this study.
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PCR errors were excluded by sequencing.
Patients included in the study were excluded by sequencing to carry a structural mutation in RPS17, RPS19 or RPS24, respectively (Primer sequences available on request).
A germline variant was excluded by sequencing normal retinal tissue.
The presence of genetic polymorphisms was excluded by sequencing the PTEN/MMAC1 cDNAs from corresponding blood mononuclear cells, which were regarded as non-neoplastic tissues of the patients where PTEN/MMAC1 cDNAs were found to be altered in the melanoma.
A possible presence of mutations within the 35S promoter or GFP sequence was excluded by sequencing of 35S-GFP cassettes obtained by PCR amplification; all sequences obtained from individual 5/6 clones 5/6a, 5/6b, 5/6h and 5/6j were identical (data not shown).
Mutations of DCX and LIS1 were excluded by sequencing or by review of brain imaging, so that patients with posterior-predominant LIS (the LIS1 pattern) had normal sequencing results for LIS1, but may or may not have had DCX testing, and vice versa.
Furthermore, because the mutation occurs downstream of all the splicing sites, the possibility of alternative splicing and/or altered pre-RNA maturations seems unlikely and this was excluded by sequencing of the cDNA of cxcr3.2 in mutants and wild types (wt).
Underquantification because of nucleotide mismatches below oligonucleotide binding sites and contamination of nested RT-PCR was excluded by sequence comparison (up to 5% nt divergence from other hCV strains, including the positive control).
Besides, classical MODY has been excluded by complete sequencing of HNF1A and GCK genes.
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