isolating and analysing nucleic acids

•  Restriction enzymes like EcoRI recognize and cut at specific palindrome sequences to generate staggered ends.
• DNA fragments separated by gel electrophoresis based on size. Electric field pulls negatively charged DNA through gel matrix.
• DNA ligase joins compatible ends. DNA polymerase fills in overhangs using dNTPs in an ATP-dependent reaction.
• Plasmids act as cloning vectors to introduce foreign DNA into bacteria for amplification. Restriction enzymes and ligase insert DNA.
• Key sequencing methods: Illumina - short fragments, fluorescent labels. SMRT - long reads, real-time detection. Nanopore - electric currents through pores.
• Sequencing enables genetic analysis, gene function/regulation, drug discovery, personalized medicine.
• Genome annotation identifies genes, regulatory regions, splicing to understand gene function.