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The size of each digested restriction fragment was estimated manually by comparison with either λ DNA concatemers (lambda ladder PFG marker, New England Biolabs) or L. lactis IL1403 SmaI restriction fragments [78], with PFGE conditions optimized for optimal resolution (pulse times of 2 to 210 s, depending on the fragment size to be determined).
Introns in the genomic sequences were identified manually by comparison to the nucleotide sequences of corresponding ESTs or cDNAs.
Alleles were sized manually by comparison with 50 350 bp size standards (LI-COR), and then scored manually from gel image files.
The models were further examined and corrected manually by comparison with AGO genes identified from other plant species using the BLASTx algorithm (http://www.ncbi.nlm.nih.gov/BLAST) [ 27].
The predicted gene models were further examined and corrected manually by comparison with AGO genes identified from other plant species using the BLASTx algorithm (http://www.ncbi.nlm.nih.gov/BLAST) [ 27].
Melt curves arising from primer-dimer amplification were identified and removed from the dataset manually by comparison with a positive control sample.
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128 clusters that fell into category 1 or 3 were manually inspected by comparison to the GenBank non-redundant (nr) database using BLAST and ψ-BLAST.
Annotations were manually refined by comparison with genes of more closely related species.
The number of terms was determined manually, often by comparison of different models.
Putative operons in Buchnera have been manually identified by comparison with the E. coli genome.
Subsequently, the annotation was manually curated by comparison to the Swiss-Prot, TrEMBL, and InterPro database [ 39, 40].
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