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Using probes from the contig, we tested for deletions within this region in DNA from 105 human tumor cell lines, predominantly derived from lung cancers.
Absence/presence of the transgenes was confirmed by Southern hybridization analysis [24] using probes from the PM3B (bp 1231 1956 as referred to the GenBank accession AY325736) and PMI (bp 271 810 as referred to the GenBank accession AAC74685) encoding region.
Fluorescence in situ hybridisation (FISH) was performed on TMAs using probes from the RP11 BACs library.
The interest in PH1 prompted the cloning of AGXT cDNA [ 23, 24], using probes from the orthologous rat gene [ 25].
BAC clones from the Sr2 locus of Hope were isolated using probes from the previously characterised 3B locus of Chinese Spring and were sequenced by a BAC-by-BAC shotgun method at 10× coverage using Applied Biosystems capillary sequencing.
To obtain high coverage in the QTL region, Agilent SureSelect (Agilent, Santa Clara, CA) target enrichment system was used to perform DNA sequence enrichment using probes from the QTL region according to genome assembly Galgal3.0.
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The construction of the contig was performed by using probes derived from the end of the starting PACs in repeated library screening.
Cross-species chromosome painting using probes derived from the rufous bettong (Aepyprymnus rufescens), the marsupial with the highest diploid number of 2 n = 32, has divided marsupial genomes into just 19 conserved segments (Rens et al. 2003).
Using probes designed from the N-terminal amino acid sequence common to the three, we isolated 14 clones of cDNA belonging to three similar sequences which probably correspond to NtEPb1 b3.
In future studies, we will use in situ hybridization to examine the mechanism of visual adaptation in A. anableps using probes designed from the opsin repertoire characterized here.
Northern blot analyses using probes generated from the full-length Atp7a cDNA revealed several specific hybridization bands, all of which were more intense in RNA samples extracted from duodenal enterocytes isolated from iron-deficient rats.
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