PCR = "Photocopier for DNA" β denature, anneal, extend
Why Taq polymerase? β Isolated from Thermus aquaticus β a bacterium living in hot springs. Heat-stable, survives the 94Β°C denaturation step. Regular DNA polymerase would denature and stop working.
Gel electrophoresis β Separates DNA fragments by size. Smaller fragments migrate farther through agarose gel toward positive electrode. DNA ladder provides size reference. Used to visualize PCR products.
Restriction enzymes β Cut DNA at specific palindromic sequences. EcoRI cuts at GAATTC. Produce sticky ends that can be joined to other DNA cut with the same enzyme. Foundation of recombinant DNA technology.
CRISPR-Cas9 β Guide RNA directs Cas9 protein to specific DNA sequence. Cas9 cuts both strands. Cell repairs by NHEJ (Non-Homologous End Joining β error-prone, disables gene) or HDR (Homology-Directed Repair β can insert new sequence). Revolutionary gene editing tool.
Why can't we make them? β Humans lack specific enzymes needed to synthesize the carbon skeletons of essential amino acids. These biosynthetic pathways were lost during evolution β easier to obtain from diet. Non-essential amino acids can be synthesized from intermediates of glycolysis and Krebs cycle.
Protein complementation β Vegetarians combine incomplete proteins to get all essential amino acids. Examples: rice + beans (rice lacks lysine, beans lack methionine β together complete), peanut butter + bread. Don't need to eat at same meal β just same day.