LAUSR

Supplemental methods The 100K Genomes Project (100KGP) is a national genome sequencing initiative approved by the HRA Committee East of England, Cambridge South (REC: 14/EE/1112). More information about this project is available online (www.genomicsengland.co.uk and https://doi.org/10.6084/m9.figshare.4530893.v5). Germline DNA samples from 78,195 individuals were sequenced using a 150bp paired-end format in a single lane of an Illumina HiSeq X instrument. Reads were aligned to the GRCh38 assembly (with decoys) using the iSAAC Aligner v03.16.02.19 and small variants were called using Starling v2.4.7. Aggregation of single-sample gVCFs were performed using gVCF genotyper v2019.02.29 (Illumina) and normalisation/decomposition was implemented by vt version 0.57721. The multisample VCF was split into 1,371 roughly equal chunks to allow faster processing and the loci of interest were queried using bcftools v1.9. In order to validate the p.(Val248Ala) variants in TUBB2A and TUBB2B, PCR reactions were performed in 25μl volumes using the Megamix (Microzone) or FastStart (Roche) kits along with the primers listed in Table S4 Thermocycling conditions included 5 minutes of initial denaturation at 95°C, followed by 30 cycles of 1 minute at 95°C, 1 minute at 58°C and 1 minute at 72°C. A final extension for 10 minutes was performed at 72°C. Prior to sequencing, PCRs reactions were purified using Exonuclease I and Shrimp Alkaline Phosphatase. Sequencing reactions were performed using BigDye v3.1 chemistry and samples were run on an ABI 3730xl/3130xl instrument.