Sepsis is a life-threatening organ dysfunction caused by a dysregulated host response to infection. As the immune system overreacts, the patient outcome can be detrimental, where every hour of delay substantially increases the mortality rate. Therefore, early diagnosis and treatment are critical for patient outcomes. The project aimed to obtain high quality and quantity bacterial DNA to be used in metagenomic sequencing for identification of pathogens and antimicrobial resistance genes, ultimately contributing to earlier diagnosis of sepsis and effective treatment. DNA extractions were performed manually and semi-robotically to compare the outcome, turnaround time, and hands-on time, considering time-sensitivity in clinical applications. Whole blood from healthy human donors were spiked with whole bacteria, bacterial DNA, and a microbial community DNA standard, where the DNA were extracted before sequencing with the MinION device. Raw sequencing data was analysed through the EPI2ME software which enabled taxonomic classification and antimicrobial resistance gene detection. Obtained results demonstrated the success of host exclusion via adaptive sampling, with high quality of data. Negative controls were expected to only contain human DNA, but the presence of different bacterial species were detected, and several unexpected species were also present throughout most DNA samples. However, expected bacterial species were still detected throughout the DNA samples, with mdf(A) gene present in two. Detection of the Escherichia coli spike-in of 10 CFU which mimics the limited amount naturally found in sepsis patients’ blood was achieved. Therefore, the project showed the potential of nanopore sequencing in detecting pathogens and antimicrobial resistance genes in blood samples, while achieving earlier sepsis diagnosis.