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Using a breast cancer dataset [40] and the average PCC across all microarray probe pairs between any two signatures, significant co-expression was identified in approximately half of the analyses when compared to 10,000 equivalent, randomly selected gene sets (empirical P values<0.05) (Fig. 4A).
Comparisons were made for all possible microarray probe pairs.
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By incorporating hairpin structures in the microarray probe, base-pairing with the additional nucleotide incorporated during labeling, and empirical melting point-determination, the platform is capable of single-nucleotide discrimination in the miRNA sequences while specifically distinguishing the mature miRNAs from longer RNAs in the total RNA sample [ 26, 31, 32].
First described by Geman and colleagues in 2004 [ 22], the TSP algorithm evaluates the relative expression of all possible pairs of genes in a microarray probe set, and selects those gene pairs for which the ordering of expression is most likely to reverse from one phenotype to the other.
Genome-wide analysis of GAS gene expression was achieved through use of a custom Affymetrix microarray that contained 16 antisense oligonucleotide probe pairs (PM + MM) for each gene in the MGAS2221 genome.
Matching of expression microarray probes to the corresponding copy number microarray probes using a 375 kb genomic window yielded 53,145 probe pairs.
The p values from Student's t-tests comparing log2 PM/MMi) with the signal differences of identical probe pairs between human and monkey microarrays were extremely small (<10-6).
An important difference between Affymetrix GeneChips and other oligonucleotide platforms is that Affymetrix uses multiple 25-mer probe pairs, while all other oligonucleotide microarrays use a single probe per gene, varying in length from 50 to 70 bases.
The Affymetrix X. laevis microarray consists of "probe sets" that are composed of 16 "probe pairs", each of which includes a 25 base pair oligo that is intended to perfectly match the target sequence.
In order to further explore this issue, microarray data for digestive glands and gills were analysed by examining sense-antisense probe pairs.
Our study showed that among the known methods for quality assessment of microarrays, the best results can be obtained using methodologies based on PM and MM probe pairs.
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