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ClustalX software (http://bips.u-strasbg.fr/fr/documentation/clustalx/ g) was used to obtain alignment with a set of KIPyV and WUPyV isolates from Italy (6, 7 ) (accession nos.

Briefly, mRNAs and ESTs were aligned to the genome using SIM4 [ 32] to obtain alignments with splice sites precisely defined.

First, all-vs-all BLASTp for all protein sequences of nine species was performed to obtain alignments with an E-value upper threshold of 10-5.

On a dataset like RV11, the improvement is much less marked (M-Coffee8 delivered alignments having an average accuracy of 37.5%) and one needs to restrict the combination to the four best non-template-based methods in order to obtain alignments with accuracy comparable to the best methods (Table 1).

Among the obtained alignments, those with TM-scores >0.6 for Fr-TM-align and those with Z-scores >2 for DaliLite were used as reference alignments, respectively.

The manually-obtained alignment with secondary structure information was used to construct a matrix for cladistic analysis based on molecular morphometrics using geometrical features and base numbering [ 16].

The highest downstream false-positive rates were obtained after alignment with Bfast, followed by CLC Bio's Beta aligner, Nextgene, Mosaik, Stampy, and Bowtie 2. Novoalign and Smalt had the lowest downstream false-positive rates followed closely by Omixon and CLC.

Regarding to the bge-miR-cand1, the precursor sequence was obtained after alignment with A. pisum genome sequences and PCR amplifications were carried out using as template cDNA of ovaries from 3-day-old female adults which had been treated with dsRNA targeting dicer-1 just after the imaginal emergence.

However, using the alignment identities in Fig. 3B(i) it can be seen that, although generally poorly conserved, the best match of 51.2% was obtained by alignment with the N2 domain.

For 88% of the transcripts, we obtained an alignment with a BAC sequence, which, excepted for five transcripts, was localized on pig chromosomes allowing 33 more transcripts to be localized on chromosomes.

Thus, the algorithm was evaluated starting from low resolution with the aim of obtaining an alignment with rotation and translation errors of less than 5° and 5 mm, respectively.

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