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Previously we identified a major cottonfiber strength QTL (qFS-c7-1) on chromosome A07 using a multi-parent advanced generation intercross (MAGIC) population. To assess the stability and transferability of this QTL and its utility in cotton breeding, we made ten new populations. These populations were devel-oped from crosses between MAGIC recombinant inbred lines, or between cotton cultivars that are differ-ent from the MAGIC parents. A total of 2801 F2 plants were grown and their fiber quality traits were measured. We also selected a subset of F3 seeds from two populations, and grew F3 progeny plots to fur-ther evaluate the stability of this QTL. Our results showed that the peak of qFS-c7-1 is at 70–72 Mb region. This QTL had a major effect on fiber strength explaining 21.9% phenotypic variance. Its effect on otherfiber quality attributes such as micronaire, short fiber content, length and uniformity varied between populations, and no effect onfiber elongation was observed. The QTL effects were stable in the popula-tions analyzed, and in different generations of the same population. The SSR and SNP markers near and within the QTL peak reported herein will assist selecting superiorfiber quality traits in breeding, with a recommendation that the parental cotton lines should be analyzed using the seven DNA markers within the QTL peak before fully implementing marker assisted selection in a cotton breeding program.