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Using stepwise deletion of the least important factor, the only parameter remaining significantly correlated with EVLWI was height (-0.11 ml/kg per cm, P = 0.001), while weight, gender and age showed no significant relationship (P = 0.65, P = 0.40, P = 0.10, respectively, in sequence of deletion).
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Deletion breakpoints were determined by Sanger sequencing of deletion PCR products and analysis by BLAST against the C. elegans genome.
Sequencing of deletions via cloning is laborious, time-consuming, prone to cloning artifacts and allows only the most abundant deletion types to be analyzed (Supplementary Notes 3 and 4).
The distribution of the discovered 40,281 PSFs across the wheat genome was inferred by comparing the EST sequences used for Affymetrix array design with the sequences of deletion-bin mapped ESTs [ 49].
The DNA sequence of the deletion in the Gria4 tm1Dgen allele is GenBank accession no.
The sequence of the deletion allele was submitted to GenBank (AY743347).
Due to the unique sequence of the deletion, we designed primers to specifically amplify the ∆90 nt mrps16 circular species using a primer that spans the junction of the deletion.
For each TP breakpoint, a global alignment between the assembled inserted sequence and the real sequence of the deletion is then performed with needle from the EMBOSS tool suite.
Breakpoints were defined as a set of coordinates on the genome spanning the genomic sequence of the deletions.
In the SR3 vs Ta and JN177 vs Ta comparisons, the GC content of the flanking sequence of the deletions was higher than that of the insertions, while the content of the indels was close to that of the insertions.
However, due to the transient approach and the analysis of genomic methylation at only a few representative sequences, subtle or highly sequence specific effects of deletion of the CXXC domain cannot be excluded.
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