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8 Cards in this Set

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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

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)

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



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



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



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





what is a vector?

. a vector is something thats used to transfer DNA into a cell


. Vectors can be plasmids or bacteriophages



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