Optimalization of DNA Extraction for Low- Biomass Microbiome Studies in Tissue from Patients with Colorectal Cancer

Abstract

Colorectal cancer is one of the most common tumors in the world, resulting in around 10% of tumor related deaths annually. Numbers of patients under 50 years old diagnosed with CRC are still rising due to unclear yet reason.

The primary objective for this study is to establish an efficient method for extracting low microbiota from tissue resected from patients with colorectal cancer, suitable for downstream application and next- generation sequencing. Initially, we aim to analyze various homogenization and bead beating conditions for the extraction of intratumoral bacterial DNA from tissue samples. Subsequently, our focus shifts to assessing the bacterial load in the samples as the second objective. This involves establishing and validating a method for the quantification of bacterial load, with a particular emphasis on developing a standard curve for accurate quantification.

The tests performed in this thesis helped us understand the importance with establishing and optimalisation of methods used for extracting of intratumoral bacterial DNA from tissue samples from patients with colorectal cancer.

Colorectal cancer is one of the most common tumors in the world, resulting in around 10% of tumor related deaths annually. Numbers of patients under 50 years old diagnosed with CRC are still rising due to unclear yet reason.

The primary objective for this study is to establish an efficient method for extracting low microbiota from tissue resected from patients with colorectal cancer, suitable for downstream application and next- generation sequencing. Initially, we aim to analyze various homogenization and bead beating conditions for the extraction of intratumoral bacterial DNA from tissue samples. Subsequently, our focus shifts to assessing the bacterial load in the samples as the second objective. This involves establishing and validating a method for the quantification of bacterial load, with a particular emphasis on developing a standard curve for accurate quantification.

The tests performed in this thesis helped us understand the importance with establishing and optimalisation of methods used for extracting of intratumoral bacterial DNA from tissue samples from patients with colorectal cancer.