Hortense makes up a reaction mix to do Sanger sequencing on a lobster. She mixes the following together: lobster DNA, primers, four DNA nucleotides (A, C, G, and T), and DNA polymerase. The reaction does not work. What was the error in Hortense's reaction?
She didn’t cycle the reaction! She has to cycle as follows: 1. 95C-to denature the double stranded DNA 2. 50-55C depends on her primers- to anneal the primers to the single stranded DNA 3. 37-42C depending on the enzyme used - DNA polymerase extends from the primers to make the new DNA.
Also, are the dNTPs special? Either radioactive or with a fluorescence to measure/detect them?
