Post-synthesis separation of metallic (m-SWNTs) and semiconducting (s-SWNTs) single-wall carbon nanotubes (SWNTs) remains a challenging process. Gel agarose chromatography is emerging as an efficient and large scale separation technique. However, the full (100%) separation has not been achieved yet, mainly due to the lack of understanding of the underlying mechanism. Here, we study the temperature effect on the SWNTs separation via gel agarose chromatography, for four different SWNT sources. Exploiting a gel agarose micro-beads filtration technique we achieve up to 70% m-SWNTs and over 90% s-SWNTs, independent of the source material. The process is temperature dependent, with yields up to 95% for s-SWNT (HiPco) at 6 °C. Temperature affects the sodium dodecyl sulfate surfactant-micelle distribution along the SWNT sidewalls, thus determining the effectiveness of the SWNTs sorting by electronic type. The sorted SWNTs are then used to fabricate transistors with very low OFF-currents (∼10−13 A), high ON/OFF current ratio (>106) and charge carriers mobility ∼ 40 cm2 V−1 s−1