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Part 4: Genomic Analysis Recombinant DNA Technology Review
  1. A enzyme binds to DNA at a specific recognition sequence and cleaves the DNA to produce restriction fragments.
  2. Most recognition sequences are , and restriction enzymes can cleave these sequences in an offset manner to produce ends.
  3. DNA fragments can be using restriction enzymes and such as plasmids and then replicated in host cells.
  4. Other that can carry larger target DNA include phages, cosmids, and artificial chromosomes (BACs).
  5. Often the cloned gene of interest should be into proteins. vectors are engineered to express large quantities of the encoded protein.
  6. Eukaryotic hosts such as cells are useful to produce eukaryotic proteins since they can perform modifications and carry very large DNA fragments.
  7. Bacterial plasmids can be used to transfer genes to plant cells in a process caled .
  8. YACs can be used to transfer genes to animal cells in a process caled direct .
  9. The chain reaction (PCR) can be used to amplify small quantities of target DNA in vitro.
  10. Genes from whole chromosomes can be cloned to form genomic . The individual chromosomes can be isolated by cytometry and pulsed-field gel electrophoresis.
  11. Libraries of transcriptionally active genes in particular cells can be constructed from DNA (cDNA).
  12. Specific clones can be recovered from a library by with a probe followed by .
  13. Specific DNA sequences can be identified with a blot and autoradiography.
  14. DNA fragments can be sequenced by chain termination. Large-scale genome sequencing can be automated by using dideoxynucleotides.
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