Genetic manipulation- genomes

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Flashcards on Genetic manipulation- genomes, created by megan.radcliffe16 on 04/04/2015.
megan.radcliffe16
Flashcards by megan.radcliffe16, updated more than 1 year ago
megan.radcliffe16
Created by megan.radcliffe16 about 9 years ago
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Outline the steps involved in genetic engineering ( recombinant gene technology). 1.Desired gene is extracted 2. gene is isolated from other DNA using electrophoresis and probes. 3. multiple copies made using PCR. 4. gene is placed in a vector. 5. vector used to insert gene into recipient. 6. recieving organism uses gene to produce desired product/characteristic.
Outline the steps involved in Electrophoresis. 1. DNA sample placed into gel wells and cover with buffer solution. 2. Battery connected. 3. Negatively charged DNA drawn toward positive electrode. 4.Shorter lengths of DNA move fastest. 5. fragments of different lengths separated.
what is a gene probe used for? used for locating specific genes to then compare the genome.
How are gene probes labelled? * attached to a specific stain. *may be a fluorescent molecule that glows in UV light. *Nucleotides can be made using 32P - shows up on photographic plates.
Outline the steps of the Polymerase Chain Reaction. (PCR) 1. mix smaple of DNA with extra DNA nucleotides and DNA polymerase. 2. Heat to 95 °C 3. Cool to 55 °C and add primers - bind to strands being replicated. 4. Raise temperature to 75 °C 5. DNA polymerase adds nucleotides to double stranded section until complete new double stranded section made. 6. Repeat.
Why do you heat to 95 °C in step 2 of the PCR? To break the hydrogen and split the double helix into single strands with exposed bases.
Why do you lower the temperature to 55°C in step 3 of the PCR? So that the primers efficiently bind to the strands being replicated.
Why do you raise the temperature to 72°C in step 4 of the PCR? It is the optimum temperature for DNA polymerase to work.
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