Studying genomes, DNA replication and Genetic Engineering Definitions

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1. Outline the steps involved in sequencing the genome of an organism: 1) the genomes are mapped 2) samples of the genomes are sheered into smaller sections (shotgun approach) 3) sections placed into BACs, transferred to E.coli for replication

  • 4) BACs taken and cultured, DNA extracted, restriction enzymes cut into smaller fragments 5) fragments sent through electrophoresis 5) sequenced using automated process 6) computers compare overlapping regions to reassemble whole BAC sequence
  • 1) pimer joins at 3' end 2) DNA polymerase attatches and adds free nucleotides 3) if a modified nucleotide added, polymerase thrown off 4) as reaction proceeds many molecules are made varying in size in each case the final nucleotide is marked
  • 3) gel immersed in a buffer solution and electric current passed through for 2 hours 4) short lengths move faster than long lengths therefore move further 5) position of fragments can be shown with a dye or radioactive marker
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2. A vector can be

  • Formed when DNA is cut using restriction enzymes, it is a short run of unpaired exposed bases seen at the end of the cut section, complimentary bases can anneal together as part of the process of recombining DNA fragments
  • Bacterial plasmid, virus, yeast cell chromosomes
  • It contains DNA that has been added to its cells as a result of genetic engineering

3. Outline the process involved in genetically engineering golden rice

  • The process of growing bacteria on agar, then transferring a replica of that growth to plates containing growth inhibitors or promoters, analysis of growth patterns gives information about genetic properties of growing bacteria
  • 1) phytoene synthase (from daffodil) and Crt 1 enzyme (soil bacteria) are inserted into the rice, near the specific promoter sequence that switches on the genes associated with endosperm development - genes expressed as endosperm (bit you eat) grows
  • 1) improving a feature of the recipient cell eg resistant crops, growth controlling gene in farms 2) engineering organisms to synthesise useful products eg insulin, golden rice

4. Electrophoresis is

  • 1) pimer joins at 3' end 2) DNA polymerase attatches and adds free nucleotides 3) if a modified nucleotide added, polymerase thrown off 4) as reaction proceeds many molecules are made varying in size in each case the final nucleotide is marked
  • 3) gel immersed in a buffer solution and electric current passed through for 2 hours 4) short lengths move faster than long lengths therefore move further 5) position of fragments can be shown with a dye or radioactive marker
  • Used to separate DNA fragments based on their size, as the negatively charged fragments are attracted to the positive electrode
  • 4) BACs taken and cultured, DNA extracted, restriction enzymes cut into smaller fragments 5) fragments sent through electrophoresis 5) sequenced using automated process 6) computers compare overlapping regions to reassemble whole BAC sequence

5. Outline PCR

  • 1) pimer joins at 3' end 2) DNA polymerase attatches and adds free nucleotides 3) if a modified nucleotide added, polymerase thrown off 4) as reaction proceeds many molecules are made varying in size in each case the final nucleotide is marked
  • 1) DNA and DNA polymerase added 2) 95C 3) hydrogen bonds break 4) primer added 5) 55C 6) DNA polymerase binds 7) 72C 8) polymerase extends section with free nucleotides
  • 4) BACs taken and cultured, DNA extracted, restriction enzymes cut into smaller fragments 5) fragments sent through electrophoresis 5) sequenced using automated process 6) computers compare overlapping regions to reassemble whole BAC sequence
  • 3) gel immersed in a buffer solution and electric current passed through for 2 hours 4) short lengths move faster than long lengths therefore move further 5) position of fragments can be shown with a dye or radioactive marker

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