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Full analysis of sequences through a stepwise variant validation procedure allowed reliable typing of 871 samples and duplicates confirmed the repeatability of the genotyping method.
Full analysis of sequences through a stepwise variant validation procedure allowed reliable typing of more than 800 great tits based on 214,357 reads; from duplicates we estimated the repeatability of typing as 0.94.
We applied a stepwise variant validation procedure to detect true alleles (Table 2); the method is broadly based on the procedures defined by Zagalska-Neubauer et al. [ 23] and Galan et al. [ 28].
In our study, sequencing one affected family member using WES, combined with stepwise variant filtering strategy, we identified mutations of collagen IV genes in 3 undiagnosed glomerulonephritis families with no extra-renal involvement.
Indeed, a stepwise variant model was assumed (with a variant rate per marker per generation of μ), and variants were given a higher probability if the individual carried an allele that was close to the ancestral allele in terms of numbers of repeats.
Using this technique we genotyped a fragment of Mhc class I exon 3 of several hundred great tits simultaneously; and applied a stepwise variant validation procedure to separate true alleles (the procedure was based on the methods defined by Zagalska-Neubauer et al. [ 23] and Galan et al. [ 28]).
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In stepwise mode, only variants with marginal p-value < 0.05 were proceeded to the next step for calculating 2nd order interactions.
At the time the LCA chip did not yet include CEP290 variants, stepwise targeted mutation analysis was performed prior to sequencing of the total coding region.
At Chr14q24, the original signal rs7141529 remained the most significantly associated variant after stepwise logistic regression; however, two further additional independent signals described by rs2189517 and rs17105852 were identified (Table 2 and Fig. 1E).
Multivariate confounding was investigated with a variant of stepwise regression, in which the stepping criterion was not a P value but the ΔB as long as it exceeded the threshold.
To identify pathogenic variants, a stepwise filtering process was used that firstly removed non-coding, synonymous and common variants (minor allele frequency, MAF >0.005 in the 1000 genome project database or our in-house database of >800 exomes).
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