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Cufflinks does not make use of existing gene annotations during assembly of transcripts, but rather assembles a minimum set of transcripts that best describe the reads in the dataset.
The Brassica unigenes were assembled using parameters that enabled the separate assembly of transcripts of paralogous genes within each diploid Brassica genome (as these differ by ~15% at the nucleotide level) but the co-assembly of transcripts of homoeologous genes (which differ by only ~3%).
Cufflinks does not make use of existing gene annotations during assembly of transcripts, but rather constructs a minimum set of transcripts that bests describe the reads in the dataset.
Further, de novo assembly of transcripts from RNA-Seq data represents a potential avenue for gene annotation [ 68– 71].
The greater sequence coverage obtained with Illumina facilitates the assembly of transcripts and enables rare transcripts to be identified.
Then, the assembly of transcripts and the quantification of transcript expression levels were based on the mapping information.
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A concern associated with de novo assembly of transcript sequences, be it Sanger derived [ 33] or 454 sequence derived [ 15] assemblies, is the contiguity of assembled sequences.
De-novo assembly of transcript sequences produced by next-generation sequencing (NGS) technologies offers a rapid approach to obtaining expressed gene sequences for non-model organisms.
As GS-FLX becomes the standard 454 pyrosequencing platform, large-scale EST sequencing and gene discovery projects will be more successful in assembly of transcript-length contigs.
De novo assembly of transcript sequences produced by next-generation sequencing technologies offers a rapid approach to obtain expressed gene sequences for non-model organisms.
De novo assembly of transcript sequences produced by short-read DNA sequencing technologies offers a rapid approach to obtain expressed gene catalogs for non-model organisms.
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