In this report we describe the characterization of a novel null HLA‐DQB1 allele, detected by polymerase chain reaction‐sequence‐based typing but not by next‐generation sequencing (NGS). The new allele, HLA‐DQB1*06:379N, was discovered in an Italian patient with acute myeloid leukemia and also in one of her healthy siblings. The new allele is largely homologous to DQB1*06:02:01:01 with a T deletion in exon 2, in codon 11, which causes a frameshift and the formation of an unusual and premature TGA STOP in codon 27. This case report underlines the importance of not removing Sanger method sequencing from routine use for high‐resolution HLA typing, but to maintain it together with NGS technology.

HLA‐A*68:02:16 differs from A*68:02:01:01 by a single synonymous nucleotide change in codon 160 (CTG>CTA).

HLA‐DPA1*01:27 differs from DPA1*01:03:01:01 by a single non‐synonymous nucleotide change in codon 15 (ACG > ATG).

HLA‐DPA1*01:86 differs from DPA1*01:03:01:05 by a single nucleotide change in codon 94 (AAG>GAG).

The new HLA‐B*51:367 differs from B*51:01:01:64 by four substitutions in exon 1.