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Primers were designed based on conserved nucleotide sequences within the 52K gene.
Based on conserved nucleotide sequences available in GenBank and specific to p25 gene (major coat protein gene) of predominant CTV isolates of India, four primer sets (CTV-F3, CTV-B3, CTV-FIP and CTV-BIP) ware designed and custom synthesized.
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A set of universal primers complementary to the ompB of Rickettsia sp. BJ-90 (GenBank Accession: AY331393) was designed using PrimerExplorer V4 software (http://primerexplorer.jp; Eiken Chemical Co., Ltd., Tokyo, Japan) based on conserved sequences determined by the nucleotide alignment of 17 ompB GenBank entries (Table 1).
The phylogenetic reconstruction was based on conserved domains.
On the other hand, presence of only 25% phenotypically positive transformants is indicative of one highly conserved nucleotide position.
In this manuscript, we devised more stringent criteria based on nucleotide conservation to extract highly conserved miRNA families.
Molecular genetic analysis of all mitochondrial tRNAs revealed a G to A transition at nt 4308; the G is a highly conserved nucleotide that participates in a GC base-pair in the T-stem of mammalian mitochondrial tRNAIle.
Assuming that regulation of miRNA expression is conserved among mammals, we used the PhastCons evolutionary conservation track that is based on nucleotide conservation among placental mammals (http://hgdownload.cse.ucsc.edu/goldenPath/hg18/phastCons44way/) to quantify evolutionary conservation of the predicted TFBSs.
Conserved residues are shaded by different intensities based on conservation level in the alignment.
Primers were designed based on highly conserved iridoviral sequences.
Such diversity confounds interpretations of prevalence data based on relatively conserved 18S rRNA sequences.
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