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We had previously applied transcript clustering to identify early molecular stages in a mouse model of glaucoma [ 39- 41].
In our laboratory, we have successfully applied transcript profiling of whole tissue as a screening tool to determine molecular changes underlying cancer, which has led to the identification of several potential markers for prostate, ovarian and pancreatic cancer (Henshall et al, 2003a, 2003b; Heinzelmann-Schwarz et al, 2004; Segara et al, 2005).
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In this study, GEMOT was applied using transcript link score cutoff = 10−95 for identifying candidate transcripts (stage II, permutation-based FDR < 0.09) and transcript causality p value cutoff = 0.005 (stage III, permutation-based FDR < 6 × 10−5).
For each dataset, we used Tophat [ 15] tool to map short reads to the hg19 reference genome and applied the transcript assembly tool Cufflinks [ 16] to estimate expressions for all transcripts with known UCSC transcript annotations [ 17].
This methodology cannot be applied for transcripts lacking SNPs in the transcript fragment analyzed (67% of the transcripts).
Currently, there are two main methods applied for transcript border analysis from bacterial RNA-seq data - manual annotation and distribution model based methods [ 7, 14].
Subsequently, SOLiD V2 fragment library protocol (Applied Biosystems) was applied and transcript ends were depleted by two rounds of Dynabeads M-280 streptavidin (Invitrogen) treatment.
32 To identify multi-gene signatures or pathways, computational pattern learning algorithms have been successfully applied to transcript or mass spectrometry profiles.
Triple match sequencing (TMS) can be applied for transcript profiling of any organism provided that annotated genomes of related species are available.
When applied to transcript levels measured in myeloid cells from 24 genotyped recombinant inbred mouse strains, we discovered that nine gene clusters are putatively modulated by a logical combination of trait loci rather than a single locus.
For these 2 markers, we applied a transcript cut-off value of 0.50 ng/μL (specificity of the cut-off is more than 90%, as confirmed in 20 healthy donor samples).
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