Aim Sanger sequencing is considered the gold standard for high-resolution HLA typing. The complexity of the HLA region leads to an inability to phase heterozygous positions thus additional testing is required to achieve allele level typing. With next generation sequencing (NGS), HLA typing should yield high-resolution typing with limited need for additional testing. Thus lowering laboratory associated costs and potentially reducing turnaround times. Methods 59 unique samples (1383 total HLA loci) previously high-resolution HLA typed were typed via NGS (HLA-A, -B, -C, -DPA, -DPB, -DQA, -DQB, -DRB1/3/4/5). 7 samples were run for inter- and intra-assay precision. Illumina TruSight HLA typing kits were used to perform the NGS testing. 24 samples were amplified per run using long-range PCR. Two technologists were utilized for library preparation. Protocols were adapted from the Illumina guidelines. Sequencing was performed using 2 × 250 bp paired end reads on the MiSeq system with a 1% spike-in of PhiX as quality control. Results 41 (2.9%) of the 1383 HLA loci had ambiguities, all within DRB1. HLA typing using TruSight HLA caused a 94.5% reduction in the number of ambiguities compared to Sanger sequencing. 2 (0.1%) of the 1383 loci were mismatched from the known Sanger sequence. Unfiltered, the overall accuracy was 99.9% and precision was 100%. Conclusions Achieving high-resolution HLA typing on 24 patients at 11 HLA loci in a single run without substantial additional testing significantly cuts the laboratory associated costs. The DRB1 ambiguities resulted from the inability to exclude rare HLA alleles due to primer binding sites within exon 2. The 2 mismatches involved 1 DRB amplicon that appears to have been contaminated. Using those mismatches, an average Q30 score of >81% (88% for DRB3/4/5) and depth of coverage ⩾ 100 was determined necessary for HLA typing assignment. Application of these criteria would have flagged the mismatched samples for repeat testing which resulted in 100% accuracy. [ABSTRACT FROM AUTHOR]