Five hundred twenty nine microsatellite markers were used to assess polymorphism at molecular level between six rice cultivars, PDKV Shriram, Heera, AC38562, Pimpudibasa, Reeta and WAB56-50 having wide variation in yield and related traits. Ninety four (17.76%) microsatellite markers showed polymorphism between these six cultivars. Maximum polymorphism was detected between Reeta and WAB56-50 (34.02%), followed by AC38562 and Pimpudibasa (31.76%), and PDKV Shriram and Heera (27.22%). The recombinant inbred line (RIL) mapping population developed from Reeta and WAB56-50 were used for assessing genetic variability based on the yield and its component traits. The analysis revealed the significant difference among the RILs for all the yield traits. Correlation and path analysis revealed the strong association of grain yield with traits like grain number, thousand grain weight, panicle length and total spikelets per panicle. Further, principal component analysis indicated 58.12% of the total variation explained by first four principal components. Therefore, these RILs could be used for mapping of QTLs associated with yield and its component traits. The polymorphic markers identified in the present study would be useful for mapping QTLs associated with yield and its component traits.