Studying genomes, DNA replication and Genetic Engineering Definitions

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1. Electrophoresis is

  • Used to separate DNA fragments based on their size, as the negatively charged fragments are attracted to the positive electrode
  • 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
  • 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
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2. Sections of DNA containing desired genes can be extracted from a donor organism using restriction enzymes:

  • It contains DNA that has been added to its cells as a result of genetic engineering
  • 1) particular r enzyme will cut (hydrolyse) DNA at a specific base sequence called a restriction site 2) sticky end 3) DNA ligase used to condense fragments together (same restriction enzyme must be used in order for complimentary sticky ends)
  • 1) the required gene is obtained 2) a copy of the gene is placed (packed and stabalised) in a vector 3) the vector carries the gene to the recipient cell 4) the recipient expresses the gene through protein synthesis
  • 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

3. A sticky end is

  • 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
  • It contains DNA that has been added to its cells as a result of genetic engineering

4. The polymerase chain reaction is basically artificial DNA replication, it can be used to generate

  • Multiple copies of a tiny sample of DNA, useful in forensics etc
  • 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

5. Germ line gene therapy involves

  • Placing the gene into embryonic cell, this technique is not currently legal and is deemed unethical
  • Placing of the gene in adult differentiated cells, examples include the placing of CFTR genes into the respiratory system cells of individuals with cystic fibrosis

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