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The Bhattacharyya distance measures the separability between two sample sequences with Gaussian distribution and is defined by B d = 1 2 ln | C i + C j | 2 | C i | 1 / 2 | C j | 1 / 2 + 1 8 ( μ i − μ j ) T C i + C j 2 − 1 ( μ i − μ j ) (7).
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For each dataset, we simulated between four and six samples sequenced at a coverage of 50X, 100X or 1000X.
No sequence differences were detected in the two pairs of replicates: between CD630 and MRSA252 and published reference sequences or between four samples sequenced both with MiSeq and earlier with HiSeq.
These two organisms are extremely closely related (perhaps strains of a single species), so we cannot distinguish between cross contamination between two samples at the sequencing stage (which is common with Illumina sequencing) and closely related copies of the gene.
Thus, the most informative comparisons look at changes in an expression ratio (involving at least two sequences) between two samples.
After the sequence tags from Solexa sequencing went through the data cleaning by removing the low quality tags (i.e., tags less than 18 nt and tags whose adaptors were null) and contaminants (adaptors and polyA), the length distribution and common/specific sequences between two samples were analyzed.
As well as defining some new VNTR sequences for the three loci (Table 3), we further noted that in some cases there were differences in the flanking regions between two samples where the VNTR locus sequence itself was identical.
Epigenetic distance corresponds to a measure of the net dissimilarity in DNA methylation profiles of a given sequence between two samples, indicating the number of pair-wise methylation differences.
A mutually subtracted RNA fingerprinting (SuRF) method has been developed that allows efficient identification of differentially expressed sequence tags between two samples.
Before comparing two samples, it is necessary to normalize the read counts to eliminate system errors and minimize sequencing depth difference between two samples.
DEGseq is a free R package to detect DE genes between two samples with or without replicates of RNA sequencing data [ 62].
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