1. Tissue Collection and Preservation: High-quality samples are essential. Tissues are often preserved using formalin-fixed paraffin-embedded (FFPE) methods or flash-freezing in liquid nitrogen. 2. Nucleic Acid Extraction: DNA or RNA is extracted from the preserved tissue. Methods such as phenol-chloroform extraction or commercial kits are commonly used. 3. Fragmentation: The extracted nucleic acids are fragmented to a suitable size for sequencing, typically through mechanical shearing or enzymatic digestion. 4. Adapter Ligation: Short DNA sequences known as adapters are ligated to the ends of the fragmented nucleic acids. These adapters are essential for the subsequent amplification and sequencing steps. 5. Amplification: The ligated fragments are amplified using polymerase chain reaction (PCR) to generate sufficient quantities of DNA for sequencing. 6. Sequencing: The prepared library is sequenced using platforms like Illumina, which provide high-throughput and high-accuracy data.
