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Specifically, we obtained 450,053 reads differentially expressed by P. pinaster infested with nematode, which assembled into 12,157 contigs; 375,168 reads for P. pinaster control, assembled into 8,808 contigs; 342,141 reads for P. pinea infested with nematode, assembled into 9,555 contigs; and 226,608 quality reads for P. pinea control, that were assembled into 4,175 contigs.
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Both raw reads and differentially expressed genes are associated to the Bioproject PRJNA287604 at NCBI.
Additionally, the reads that differentially aligned between two parental genomes (aligned to only one of the genomes or with a different number of mismatches) were considered as allele-specific reads.
DBA: Diamond Blackfan anemia; Rpl11: Ribosomal protein L11; HSC: Hematopoietic stem cells; FPKM: The number of fragments per kilobase of transcript per million mapped reads; DEGs: Differentially expressed genes; RPs: Ribosomal proteins.
A read that maps to both genes equally well should be discarded, because if it were counted for both genes, the extra reads from the differentially expressed gene may cause the other gene to be wrongly called differentially expressed, too.
We report allelic imbalance as: the proportion of reads that are differentially expressed ((allele1 - allele2)/(allele1 + allele2)).
The mapped reads point to differentially methylated areas across the genome, with some marked preferences for hypermethylation or hypomethylation in certain genomic regions.
As an example, sequencing reads for the differentially activated miRNA, miR-422a are shown in Table 3 with a complete list of miRNAs in Table 4.
We also removed all reads that would differentially map to one parental sequence and not to the other if one of the reference sequences contained one or more 'N's for this read.
The DiffKAP pipeline analyses a pairwise dataset by identifying differentially expressed k-mers (DEKs) first, then obtaining differentially expressed reads (DERs) and finally annotating the DERs to gather differentially expressed genes (DEGs).
The differentially expressed reads were mapped against the non-redundant set of nucleotide sequences at NCBI (http://www.ncbi.nlm.nih.gov/nucleotide/) and miRBase.
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