1 / 37

TECHNIQUES IN BIOTECHNOLOGY

TECHNIQUES IN BIOTECHNOLOGY. DNA EXTRACTION. http://learn.genetics.utah.edu/content/labs/extraction/. Purpose: to purify a DNA sample Uses genetic testing crime scene investigation gene studies. Steps: . collect cells remove DNA from cell purify it use it!. Ex. Insert for plasmid.

caia
Download Presentation

TECHNIQUES IN BIOTECHNOLOGY

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. TECHNIQUES IN BIOTECHNOLOGY

  2. DNA EXTRACTION • http://learn.genetics.utah.edu/content/labs/extraction/

  3. Purpose: to purify a DNA sample • Uses • genetic testing • crime scene investigation • gene studies

  4. Steps: • collect cells • remove DNA from cell • purify it • use it!

  5. Ex. Insert for plasmid • From someone else’s DNA • ex. fish gene in strawberries, • jellyfish gene in plants

  6. In order to do these things, we need a way to make many copies of the genes we want

  7. Making more DNA: PCR • purpose: to make many copies of a specific piece of DNA Steps: • denature DNA using temperature • add in the ingredients (ATGC, enzymes, primers) to copy it • repeat

  8. Making an insert: Polymerase Chain Reaction

  9. Uses • make the desired gene/piece of DNA to use it to make proteins, study it, etc.

  10. To Make a Recombinant Plasmid: Insert • Cut the plasmid and the insert with the same restriction endonuclease to make complementary sticky ends. • Combine the sticky ends using ligase. • ligase: enzyme used to join DNA together 3. Introduce the recombinant plasmid into bacteria.

  11. Making a Recombinant Plasmid

  12. - - - - - - - - + - + - - - - ++ -++ - - ++ - ++ + + + + + - - - - - - - - - - - - + + + + + Bacterial Transformation • introduction of foreign DNA into a bacterial cell • plasmid is used as a vector, a vehicle by which DNA can be introduced into host cell phospholipid bilayer Ca2+ ions plasmid

  13. Following transformation bacteria are grown in medium with antibiotic… Only the bacteria that have the plasmid (and therefore the antibiotic resistance) will survive.

  14. Example plasmid: Origin of Replication: • where the plasmid starts to duplicate itself • the specific sequence MUST NOT be cut by restriction endonucleases or it won’t be able to replicate

  15. Using Bacteria as Production Factories • easy to grow • no ethical issues • small genome • easy to manipulate

  16. DNA SEQUENCING • purpose: find out the order of DNA, the “fingerprint” • uses: forensics, genetic testing, studying mutations

  17. Steps • like PCR, put your template (DNA you want to “read”) with the nucleotides, primers, enzymes, and use heat to make “more” • some of the nucleotides have fluorescent labels and stop the chain from being made

  18. results in a soup of different-length fragments • separate them by size, and “read”

  19. Gel Electrophoresis • used to “see” the DNA • used in sequencing and to determine the origin of DNA ex. Tsunami baby

  20. Sequencing

  21. Ex. RFLP: Restriction Fragment Length Polymorphism Comparison of different lengths of DNA fragments produced by restriction enzymes to determine genetic differences between individuals

  22. Steps

  23. CLONING http://learn.genetics.utah.edu/content/tech/cloning/

  24. Therapeutic cloning • used to produce tissue that is identical to the donor, to prevent rejection

  25. Reproductive Cloning • creates an organism with the same genetic material (DNA) as the original organism – an EXACT COPY of the donor

  26. Dolly the Sheep • the first cloned sheep

  27. Common uses of biotechnology: • Making "stuff” • proteins, enzymes, medication, etc. can be produced by engineered bacteria! • Food can be altered to have new traits • Cloning (therapeutic and reproductive) • Genetic screening • crime cases, relationship, genetic screening, etc. 3. Gene Therapy

  28. Gene therapy • desired gene is inserted into cell's nucleus using a retrovirus as a carrier • defective gene replaced by functional gene

  29. Ex. ADA deficiency • adenosine deaminase deficiency • little immunity with low chances of recovery - the T-cells of a four-year-old were removed, modified and re-inserted to fix her immune system

More Related