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The corresponding chimpanzee cDNA and protein sequences were extracted and compared to the human ortholog by using the EMBOSS sequence analysis package.
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We aligned these protein sequences with human protein sequences using the Emboss Needle program (which implements the Needleman-Wunsch algorithm [ 27]) to extract identity, similarity and gap values for each rhesus protein model.
Intron/exon boundaries were identified by alignment of clover or legume orthologue cDNA with clover genomic sequence orthologues, or publicly available genomic legume databases such as M. truncatula or Lotus japonicus genomic sequence using the EMBOSS (European Molecular Biology Open Software Suite) est2genome programme.
The GTS1 protein sequences from both Oryza sativa and Zea mays were aligned to the Arabidopsis thaliana sequence using the EMBOSS global pairwise alignment algorithm [ 12] to get the percent identity between the proteins.
Introns were assigned by aligning the unigenes with the melon genomic sequence using the Emboss: est2genome [ 105].
ORFs were identified by using the EMBOSS software (version 5.0.0; emboss.sourceforge.net).net
Nucleotide sequence identities were calculated after the pairwise sequence alignment using the EMBOSS Water program (http://www.ebi.ac.uk/Tools/psa/emboss_water/nucleotide.html ).html
Sequence analysis was performed using the Emboss software suite [ 37], Clustal W [ 38] and 3DCoffee [ 39].
Sequence manipulation was carried out using the EMBOSS suite of programmes [ 54].
The sites were mapped to 3 kb of promoter sequence of the β-catenin target genes using the Emboss program fuzznuc [31] allowing for 1 mismatch.
Primer pairs were matched against bovine sequence from GenBank, NT and BTGI databases using the EMBOSS [ 26] primersearch tool.
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