GENE 320 Lecture Notes - Lecture 21: Genbank, Sanger Sequencing, Protein Structure
Document Summary
Steps are repeated over and over using thermocycler to amplify dna exponentially. Some info about nucleotide sequence of target dna is required to synthesize primer. Minor contamination from other sources can cause problems. Allele speci c probes for genetic testing can be synthesized. Reverse transcription pcr helps study gene expression used to generate ds-cdna. Real time pcr allows researchers to quantify ampli cation reactions as they occur in real time. Created by cutting dna with different restriction enzymes and separating dna fragments by gel electrophoresis, which separates fragments by size. Fragments visualized on gel by staining with ethidium bromide and illuminated by uv light establishes number of, order of, and distances between restriction enzyme cleavage sites on cloned segment of dna. Used to identify which clones in library contain given dna sequence. Used to characterize size fragments from restriction digest. Used to determine whether gene is actively being expressed in given cell or tissue.
