In this article, the authors provide an in-depth protocol for detecting somatic mutations from single cells. Current methods for analyzing these cancer-causing mutations involve clonal expansion of cells and bulk sequencing. However, somatic mutations rarely amplify throughout the clone and are unique to each single cell, thus making these mutations difficult to identify using bulk sequencing. To sequence at the single cell level, Whole Genome Amplification (WGA) must be performed, however this often introduces artificial variations in the genome. To improve the accuracy and efficiency of single cell sequencing, the authors developed a technique called Single Cell Multiple Displacement Amplification (SCMDA). This novel method involves isolating single cells or nuclei with Cell Microsystems CellRaft AIR System and microwell array technology. SCMDA performed on these single cells or nuclei allows for reliable amplification of the whole genome while SCcaller software filters out any remaining artifacts from the amplification process. Therefore, by following the detailed protocol provided, researchers can discover mutations in single cells with high accuracy and fewer processing steps than current methods.
