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Isolated PAC clones were completely digested with BamHI and subcloned into plasmid vector.
Twelve BAC clones were completely sequenced.
In addition, 35 187 FL-cDNA clones were completely sequenced.
Following this, PCR products were electroporated into AbleK bacterial cells (Stratagene) and selected clones were completely sequenced.
In addition, for the Paris strain five fosmid clones were completely sequenced by transposon-assisted sequencing (Finnzymes, TGS II kit).
The collection has eventually expanded to 578K FL-cDNA clones, among which 35K cDNA clones were completely sequenced and annotated (available at Knowledge-based Oryza Molecular biological Encyclopedia (KOME): http://cdna01.dna.affrc.go.jp/cDNA/).jp/cDNA/
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The function of 9 of these clones was completely unknown.
Interestingly, differences in methylation largely stem from allelic methylation pattern, whereby some clones are completely unmethylated.
Within each of the two B. atticus clusters, clones are completely intermingled, without any hint of geographical or population trend.
Intriguingly, the structure of the resulting adult eyes that had lost the PTEN clones was completely normal, indicating that the surrounding tissue was able to compensate for the loss of the clones.
Furthermore, a N. gruberi priS cDNA clone was completely sequenced (see Additional File 1 for complete listing of the datasets).
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