Thailand is the world leader in the production of latex extracted from the rubber tree (Hevea brasiliensis). However, the most cultivated clone RRIM 600, is highly susceptible to diseases, and there is economic incentive to develop new rubber tree clones. Four rubber tree clones (T2, SK1, NK1 and SK3) that have high latex yield potential from plantations in Southern Thailand were selected for this study. Yield performance, latex biochemical parameters and anatomical characteristics of bark were monitored for two years, using RRIM 600 clones in the same fields as paired controls. The average yields of the clones SK1, NK1 and SK3 were 129.3, 74.2 and 53.9 g per tree per tapping, respectively, surpassing the paired RRIM 600 controls (94.3, 49.9 and 43.9 g per tree per tapping in matching order). There was a difference in girth increment of SK1, SK3 and T2 clones when compared with RRIM 600, whereas the clones SK1 and T2 had higher renewed bark thickness than the paired RRIM 600. The anatomical measurements showed that the diameter of the latex vessels and density of latex vessels mm−2 were the highest in clone NK1, which also had the best latex biochemical parameters. This indicates NK1 is superior, and supports its use in Hevea breeding programs to improve latex yield. Our genetic characterization and assessment of the four clones selected used Random Amplified Polymorphic DNA (RAPD) and Simple Sequence Repeats (SSR). Seventeen recommended rubber clones were included as references. The clones SK3 and SK1 were closely related to RRIM 600 with similarity coefficients of 0.891 and 0.809, while NK1 and T2 were closely related to RRIT 250 (0.836) and RRIC 110 (0.864), respectively.