1 / 33

A Lot More Advanced Biotechnology Tools

A Lot More Advanced Biotechnology Tools. DNA Sequencing. DNA Sequencing. dideoxynucleotides ddATP, ddGTP, ddTTP, ddCTP missing O for bonding of next nucleotide terminates the growing chain. 2. 1. 3. 4. 2. DNA Sequencing. Sanger method synthesize complementary DNA strand in vitro

juan
Download Presentation

A Lot More Advanced Biotechnology Tools

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript


  1. A Lot More Advanced Biotechnology Tools • DNA Sequencing

  2. DNA Sequencing • dideoxynucleotides • ddATP, ddGTP, ddTTP, ddCTP • missing O for bonding of nextnucleotide • terminates the growing chain

  3. 2 1 3 4 2 DNA Sequencing • Sanger method • synthesize complementary DNA strand in vitro • in each tube: • “normal” N-bases • dideoxy N-bases • ddA, ddC, ddG, ddT • DNA polymerase • primer • buffers & salt

  4. Reading the sequence • Load gel with sequences from ddA, ddT, ddC, ddG in separate lanes • read lanes manually & carefully • polyacrylamide gel

  5. Fred Sanger 1978 | 1980 • This was his 2nd Nobel Prize!! • 1st was in 1958 for the structure of insulin

  6. The Sanger Method: DNAi video tutorial

  7. Advancements in sequencing • Fluorescent tagging • no more radioactivity • all 4 bases in 1 lane • each base a different color • Automated reading

  8. More Advancements in sequencing • Capillary tube electrophoresis • no more pouring gels • higher capacity & faster Applied Biosystems, Inc (ABI) built an industry on these machines

  9. Big labs! • economy of scale • PUBLIC • Joint Genome Institute (DOE) • MIT • Washington University of St. Louis • Baylor College of Medicine • Sanger Center (UK) • PRIVATE • Celera Genomics

  10. Automated Sequencing machines • Really BIG labs!

  11. Human Genome Project • U.S government project • begun in 1990 • estimated to be a 15 year project • DOE & NIH • initiated by Jim Watson • led by Francis Collins • goal was to sequence entire human genome • 3 billion base pairs • Celera Genomics • Craig Venter challenged gov’t • would do it faster, cheaper • private company

  12. Different approaches Craig Venter’s method gov’t method “map-based method” “shotgun method” 1. Cut DNA entire chromosome into small fragments and clone. 2. Sequence each segment & arrange based on overlapping nucleotide sequences. • Cut DNA segment into fragments, arrange based on overlapping nucleotide sequences, and clone fragments. • 2. Cut and clone into smaller fragments. 3. Assemble DNA sequence using overlapping sequences.

  13. Shotgun!

  14. Human Genome Project • On June 26, 2001, HGP published the “working draft” of the DNA sequence of the human genome (4 years ahead of schedule). • Historic Event! • blueprint of a human • the potential to change science & medicine

  15. Sequence of 46 Human Chromosomes 3G of data 3 billion base pairs

  16. Things Are Strange In Here:

  17. Raw genome data

  18. NCBI GenBank • Database of genetic sequences gathered from research • Publicly available on Web!

  19. Maps of human genes… • Where the genes are… • mapping genes & their mutant alleles

  20. gene gene polypeptide 1 Defining a gene… protein RNA gene polypeptide 2 polypeptide 3 • “Defining a gene is problematic because… one gene can code for several protein products, some genes code only for RNA, two genes can overlap, and there are many other complications.” • – Elizabeth Pennisi, Science 2003

  21. Exons (regions of genes coding for protein, rRNA, tRNA) (1.5%) Types of DNA sequences in the human genome Repetitive DNA that includes transposable elements and related sequences (44%) Introns and regulatory sequences (24%) Unique noncoding DNA (15%) Repetitive DNA unrelated to transposable elements (about 15%) Alu elements (10%) Simple sequence DNA (3%) Large-segment duplications (5–6%)

  22. And we didn’t stop there…

  23. Genome Sizes and Estimated Numbers of Genes*

  24. What have we found? • When you go looking…

  25. …you will certainly find something!

  26. Sing A Silly Song Extolling Automation!

  27. Ethical Questions…

  28. 1. You have a familiar history of a terminal genetic disease. A genetic test exists for the disease. Would you take the test to see if you will have the disease?

  29. 2. As a condition of your continued employment, your boss wants you to have a genetic screening test and the results sent to the company. Will you get the test?

  30. You and your spouse have decided to have a child. You have the resources to pre-determine your child’s gender. Would you do this? • What about your child’s intelligence?

  31. 5. You discover your favorite food contains genetically modified ingredients. Will you continue to eat the food?

  32. As part of a routine medical procedure, your doctor discovers that you have a rare, beneficial variant of a protein that protects you from heart disease. Should your doctor be able to patent the protein? • Should you be entitled to any money from the patent?

More Related