Which of the following methodologies was not used in Human Genome Project?

DNA fingerprinting is not a methodology involved in the Human Genome Project.

Expressed sequence tags and sequence annotation are two major methodologies adopted in the Human Genome Project (HGP). HGP also involved producing genetic and physical maps of chromosomes. A genome map describes the order of genes or other markers and the spacing between them on each chromosome. DNA sequencing is the process of determining the order of nucleotides in DNA. For sequencing, the total DNA from a cell is isolated and converted into random fragments of relatively smaller sizes, cloned, and sequenced using automated DNA sequencers that worked on the principle of a method developed by Frederick Sanger.

Mapping involves dividing the chromosomes into smaller fragments and ordering (mapping) them to correspond to their respective locations on the chromosomes.