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8 Cards in this Set
- Front
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what is gene technology? |
. the polymerase chain reaction - produced lots of copies of an identical gene . In vivo and in vitro gene cloning . DNA probing - used to identify genes |
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how does using reverse transcriptase produce DNA fragments? |
. mRNA molecules are easy to obtain . used a templates to make lots of DNA . enzyme Reverse transcriptase makes DNA from a RNA template . DNA produced is called cDNA (complimentary DNA) |
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how does using the polymerase chain reaction produce DNA fragments? |
. reaction mixture is made up of, SNA sample, free nucleotides, primers and DNA polymerase . DNA is heated to 95oC to break hydrogen bonds between base pairs . mixture is then cooled to 60oC so primers can bind . reaction mixture is heated to 72oC so DNA polymerase can work . lines free nucleotides to their complimentary base pairs . two new copies of fragment is formed and one cycle of PCR is complete . the multiplication of DNA when PCR is done over and over again is exponential |
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hw does using restriction endonuclease produce DNA fragments? |
. palindromic sequences of DNA are DNA that read the same in opposite directions . these are known as recognition sequences . restriction endonuclease cut at these places . different restriction end nucleases cut at different palindromic sequences . DNA sample is incubated with specific restriction endonuclease's . sometimes the cut leaves sticky ends . this makes binding DNA to other bits of DNA easier |
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what is the difference between in vivo cloning and in vitro cloning? |
. in vitro cloning is where the gene is copied over and over by using PCR . in vivo cloning is where the gene is copies are made within a living organism |
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how do you create recombinant DNA for use in in vivo cloning? |
. Vector DNA (not yet containing the target gene) is isolated . cut open using restriction endonuclease's. . sticky ends of vector DNA are complimentary to sticky ends of DNA fragment containing the gene . vector DNA and DNA fragment are mixed together with DNA ligase which joins sticky ends of both together (ligation) . this is called recombinant DNA |
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what is a vector? |
. a vector is something thats used to transfer DNA into a cell . Vectors can be plasmids or bacteriophages |
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How do you transform cells for use in in vivo cloning? |
. recombinant DNA is used to transfer a gene into cells called host cells . these host cells that have taken up the gene are said to be transformed . heat shocks encourage bacteria to take up these plasmids . with a bacteriophage vector, the bacteriophage will infect the host bacterium by injecting its DNA into it |
