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We used 161 independently collected samples for replication.
We selected 10% of the samples for replication, and the results were 100% concordant.
This limitation has resulted in small sample size of discovery stage and a difficulty of collecting samples for replication analysis.
It is crucial, therefore, to follow up any positive signals of association with genotyping in independent samples for replication.
Four anonymous genomic DNA samples for replication sequencing were provided by the Estonian Genome Center of the University of Tartu EGCC).
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In November 2008, Bertram et al. [ 28] reported on a GWAS analyzing 500,000 SNPs in a sample of 410 families and using multiple other samples for replications.
To increase the power to detect association, the larger sample was used as a discovery sample and the smaller sample was used as an independent sample for replication.
The polygenic architecture of gF and gC was previously confirmed using the Cognitive Aging Genetics in England and Scotland (CAGES) cohort as the discovery sample and the Norwegian Cognitive NeuroGenetics (NCNG) adult lifespan sample for replication (Davies et al. 2011).
The case control replication of the SVEP1 association in the Dutch did not reach statistical significance indicating that the initial association was spurious or that there is a population stratification issue in the samples used for replication.
All samples for the replication study were collected with the same protocol using EDTA anticoagulant (Table 1).
Thus, the actual required sample size for replication is even larger than the one estimated based on the effect size reported in the initial discovery study.
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