Use policyThe full-text may be used and/or reproduced, and given to third parties in any format or medium, without prior permission or charge, for personal research or study, educational, or not-for-pro t purposes provided that:• a full bibliographic reference is made to the original source • a link is made to the metadata record in DRO • the full-text is not changed in any way The full-text must not be sold in any format or medium without the formal permission of the copyright holders.Please consult the full DRO policy for further details. We report a refined determination of muonium 1S-2S transition frequency and its isotope shift with deuterium by recalibrating the iodine reference lines using an optical frequency comb. The reference lines for the muonium and deuterium 1S-2S transitions are determined with a precision of 2.4 × 10 −10 and 1.7 × 10 −10 , respectively. An updated muonium-deuterium 1S-2S isotope-shift frequency is derived from these references to be 11 203 464.9(9.2)(4.0) MHz, in agreement with an updated bound-state quantum-electrodynamics prediction based on 2010 adjustments of Committee on Data for Science and Technology and 2.3 times better in the systematic uncertainty than V. Meyer et al.