A 2-yr field study was conducted during 2007 and 2008 at Stoneville, MS, to determine the effect of twin-row (two rows 38 cm apart on 102-cm beds) and single-row (on 102-cm beds) patterns and glyphosate POST applications with and without fluometuron S-metolachlor PRE on cotton canopy closure, weed control, and lint yield in two cultivars (‘DP117B2RF’, early maturity, hairy leaf; ‘DP164B2RF’, mid to full maturity, smooth leaf) under an irrigated environment. The experiment was conducted in a split–split plot arrangement of treatments in a randomized complete block design with row pattern as the main plot, cultivars as the subplot, and herbicide programs as the subsubplot. Cotton canopy closed 2 wk earlier in the twin-row pattern compared to the single-row pattern. Canopy closure was unaffected by cultivars and herbicide programs. Control of nine predominant weeds was sufficient (≥ 95%) to support cotton production. Total weed dry biomass was reduced by 35% in twin rows compared to the single-row pattern, 15% in DP117B2RF compared to DP164B2RF cultivar, and ≥ 97% with glyphosate early POST (EPOST), EPOST followed by (fb) mid-season POST (MPOST), EPOST fb MPOST fb late POST (LPOST) following PRE herbicides or three applications of glyphosate POST only without PRE herbicides compared to no herbicide. Cotton grown in twin-row pattern produced 6% higher lint yield than single-row cotton. Cultivar DP117B2RF produced 23% higher lint yield than cultivar DP164B2RF. Lint yields were higher with glyphosate EPOST fb MPOST, EPOST fb MPOST fb LPOST following PRE herbicides or three applications of glyphosate POST only without PRE herbicides (1,210 to 1,230 kg/ha) compared to glyphosate EPOST following PRE herbicides (1,130 kg/ha). These results demonstrated that cotton grown in twin-rows closed canopy early and produced higher lint yields than cotton grown in single-rows.