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Alignment of transcript regions to genomic sequences provided a preliminary exon map of each gene.
For example, alignment of transcript I displayed six exons and five introns, which is identical to the gene structure predicted by the EST evidence.
The alignment of transcript sequences to genomes is currently the most effective way to detect alterations of mature mRNA at a large scale.
We used GenomeThreader [ 61] transcript alignments in exchange of PASA's standard GMAP alignments [ 62] as GenomeThreader supports alignment of transcript evidence to multiple (nearly identical) loci.
In contrast with the majority of other tools for the analysis of alternative splicing, ASPic performs a multiple alignment of transcript data to the genomic sequence and refines exon-intron boundary alignments through dynamic programming [ 27].
MapNext provides four main analyses: (i) unspliced alignment and clustering of reads, (ii) spliced alignment of transcript reads over intron boundaries, (iii) SNP detection and estimation of minor allele frequency from population sequences, and (iv) storage of result data in a database to make it available for more flexible queries and for further analyses.
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Alignment of transcripts in the cases studied was performed with the ClustalW algorithm in the MEGA v.5.05 tool package [ 43], using by default parameters.
Alignment of transcripts to the genome indicated that 16733 and 16969 genes were expressed in the control and cisplatin-treated samples, respectively.
Criteria for presence of exon 1 included: comparison of the number and location of exons in orthologous genes, alignment of transcripts of the gene as reported by different databases, and alignment of the 5' end of the transcript with the putative start site and signals in the gene.
The transcripts were aligned on the genome using PASA [ 42], an eukaryotic genome annotation tool that uses spliced alignments of transcript sequences to automatically model and update gene structures.
Finally, our analyses show that the low conservation observed in cross species alignments of transcript exonic structures between human and mouse [ 33] is intimately related to the evolution of the exon sequences and the regulators of splicing.
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