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When necessary, confirmation of the disrupted sequence was achieved by PCR.
Besides the single mutation, the fragment encoded an otherwise wildtype sequence and was intended to replace the disrupted sequence at the native locus.
Genomic DNA from mice was amplified by PCR using the primers: 5-GTCATTTCCTGACCTCTGACC-3 and 5-AACCGCATTATTCCATGGGGC-3 to detect WT DNA and primers 5-AACCGCATTATTCCATGGGGC-3 and 5-GCCTGCTCTTTACTGAAGGCTC-3 to detect the disrupted sequence.
Using a custom SNP microarray we validated 150 SPGs in a cohort of 468 individuals, with every individual genome averaging 36 disrupted sequence variations, 15 in homozygote form.
As for the detection for progenies of GD individuals, which might contain homologous disrupted sequence, a heteroduplex formation procedure after mixing with WT PCR product is necessary to avoid omission.
Similar(55)
TALENs with cognate sequences in the S or C open-reading frames (ORFs) efficiently disrupted sequences at the intended sites and suppressed markers of viral replication.
For example, if many post-integrationally disrupted sequences are present, or sequences with a variable amount of inserts, they can destroy the alignment.
The genomic arrangement of XacFhaB and XacFhaC points to a situation where downstream genes of XacFhaB have disrupted sequences probably preventing the making of functional proteins.
If contigs with 5× 10× coverages were included, these numbers considerably elevate (15 sequences with homopolymer run associated errors and 9 disrupted sequences outside the homopolymer region).
Comparison of the coverage read numbers of OR sequences originated from genomic DNA versus cDNA indicated that 22 lizard OR genes (19 intact and 3 disrupted sequences) were not detected for their expression in the olfactory epithelium (fig. 3).
Some TvBspA genes are potential pseudogenes with 98 entries currently being annotated as such as they possess ORF disrupting sequence features.
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