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Biotechnology Introduction Tools Process Applications Biotechnology Introduction – basic idea – Gene is identified and excised from one organism – Gene is placed in vector (plasmid) and amplified – Gene is transferred to new organism or used in host organism to make protein product Biotechnology - Tools Restriction endonucleases – Nucleases cut nucleic acid – at first seem non specific – Linn & Abner discover that some strains of bacteria are able to resist bacteriophage infection by digesting infecting DNA – Different bacteria produce different restriction enzymes Biotechnology - Tools Restriction Endonucleases – Cut at specific 4, 6, or 8 base sites – Site is a “palindrome” » Racecar » Madam I’m Adam » Damit I’m Mad – Some restriction enzymes generate “sticky ends” Biotechnology - Tools Biotechnology - Tools Plasmids – Carry an antibiotic resistance marker – Carry restriction sites in convenient locations to insert DNA – Carry characteristics that allow the plasmid to reproduce in several organisms Biotechnology - Tools Biotechnology - Tools Polymerase Chain Reaction (PCR) – Allows any segment of DNA to be amplified chemically in minutes – Uses a thermostable DNA polymerase – Machine can cycle every 60-90 seconds Biotechnology - Tools Biotechnology - Tools Agarose Gel Electrophoresis – Can separate DNA fragments made with restriction enzymes – Can separate PCR made DNA – Can be used to sequence DNA Biotechnology - Tools Biotechnology - Process Basic process worked out by Cohen & Boyer Cut plasmid DNA and target DNA with same restriction enzyme Mix DNA and allow sticky ends to match up Select DNA clones having target gene Biotechnology - Tools