PCR, gel electrophoresis, cutting out DNA fragments using restricion enzymes.
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how does PCR work?
you collect the sample you want and the PCR will amplify it. you cut out the DNA section you want and put it ina mixture with free nucleotides, DNA polymerase and primers.
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what are primers?
short pieces of DNA that are complementry to the bases at the start of the section you want.
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then what happens with the mixture? stage 1
it is heated to about 94-96'C, this breaks the hydrogen bonds between chsins and so now there are 2 seperate chains. then the mixture is cooled to 50-60'C which allows the primers to attach to each 3' end of each strand.
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then what happens with the mixture? stage 2
it is then heated to 72'C for DNA polymerasse to attach to the nucleotides, DNA polymerase then replicates the region of DNA and you have 4 strands.
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what happens in electrophoresis? part 1
it seperates out DNA fragments. you make wells in the gell and add in DNA strands in.
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what happens in electrophoresis? part 2
you then connect the box to a power supply so the DNA fragments move towrds the positive electrode, then you stop the experiment..
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what happens in electrophoresis? part 3
you shine a flouresent light on them and they should show up.
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what do restriction enzymes do?
they are used to cut out DNA fragments
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what is gentic engeneering?
the manipulation of an organisms DNA
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what is it used for?
to make something that we want or to improve certain features.
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how is it caried out? stage 1
the required gene is obtained, a copy of the gene is placed into a vector, the vector carries the gene to the recipient cell, the recipient expresses the gene through protein synthesis.
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what are then new combination of bases in the DNA called?
recombinant DNA
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what is pharming?
pharmaceuticals are produced using genetically modififed organisms, such as animals.
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what is gene therapy?
it involves altering alleles inside cells to cure genetic disorders.
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what are the 2 types of gene therapy?
somatic and germ line.
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what is somatic therapy?
involves alternating the alleles in the body cells, particularlly the most affected by the disorder. it targets specific cells and doesnt affect the sex cells so the dissorder could still be inherited.
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what is germ line therapy?
involves altering the allleles in the sex cells, meaning that every cell of any offspring wouldnt have the disease, but currently illegal.
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how is DNA sequencing carried out stage 1
uses a similar mixture to PCR, you add in DNA nuclotides that have flouresent markers on them.
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how is DNA sequencing carried out stage 2
the modified nuceotides 'terminator bases' throw DNApolymerase off and stop the chain growing. so you end up with different lenghts of DNA
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how is DNA sequencing carried out stage 3
each base has its own colour, all the strands are ran through a mchine and a laser reads the colour sequence-starting with the shortest strands.
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Other cards in this set
Card 2
Front
how does PCR work?
Back
you collect the sample you want and the PCR will amplify it. you cut out the DNA section you want and put it ina mixture with free nucleotides, DNA polymerase and primers.
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