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Moerover multiple blasts in the reference citrus whole genome sequence (http://www.phytozome.net/clementine.php) of the corresponding sequences comfort this hypothesis (data not shown).
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The identified thresholds reflected the median similarity of SSU and LSU ribo-tags (98% and 93% respectively; see Figures S1 and S2 in SI) to their best BLAST match in the reference databases.
The annotations were verified with BLAST searches of features in the reference sequence [ 15].
To identify the putative function of catfish transcripts, all the sequences were blasted against the reference proteins available in NCBI RefSeq and Uniprot/Swiss-Prot protein databases using BLASTX with E-value ≤ 1e-10.
Firstly, for the in silico analysis, the primer sequences were blasted against the reference 'Golden Delicious' apple genome and the NCBI database.
In case of a negative blast result, an additional nucleotide blast with the reference coding sequence was performed to exclude false negative results.
We adopted a conservative approach and selected only these transfrags to blast against the reference database.
To determine the exon intron structure of the pJAB1 gene, we performed a blast search in the pig reference genomic sequence database (Sscrofa10.2) using the pJAB1 cDNA sequence.
As shown in Table 5, BLAST analysis of the reference assembly and de novo assembly contigs revealed that 24,440 unique Uniprot accessions were represented, suggesting that the expressed short reads provided good coverage of the catfish transcriptome.
The regions corresponding to the LTR and HTLV-1 genome were subjected to a blast search against the reference sequences described in Additional file 1: Table S1.
Transcripts can then be annotated by blasting against a reference genome but those assembled transcripts which are not present in the reference genome, such as those unique to an accession, are also preserved and can be annotated independently.
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