![Page 1: Biotechnology Techniques How to make Recombinant DNA Gel Electrophoresis PCR Summarize: What is this technique? Draw and label a diagram to show this technique](https://reader030.vdocuments.us/reader030/viewer/2022032802/56649e0c5503460f94af5727/html5/thumbnails/1.jpg)
Biotechnology TechniquesHow to make
Recombinant DNA
Gel Electrophores
isPCR
Summarize: What is this technique?
Draw and label a diagram to show this technique
What are the main tools or
materials involved?
Applications: What is this
being used for?
![Page 2: Biotechnology Techniques How to make Recombinant DNA Gel Electrophoresis PCR Summarize: What is this technique? Draw and label a diagram to show this technique](https://reader030.vdocuments.us/reader030/viewer/2022032802/56649e0c5503460f94af5727/html5/thumbnails/2.jpg)
Chapter 20: Terms to Know
1. Genetic engineering
2. Biotechnology
3. Recombinant DNA
4. Gene cloning
5. Restriction enzymes
6. Sticky ends
7. DNA ligase
8. Cloning vector
9. Nucleic acid hybridization
10. Genomic library
11. cDNA library
12. PCR
13. Gel electrophoresis
14. Southern blotting
15. DNA microarray assays
16. SNPs
17. RFLPs
18. Stem cells
19. Gene therapy
20. GMO (genetically modified organism)
![Page 3: Biotechnology Techniques How to make Recombinant DNA Gel Electrophoresis PCR Summarize: What is this technique? Draw and label a diagram to show this technique](https://reader030.vdocuments.us/reader030/viewer/2022032802/56649e0c5503460f94af5727/html5/thumbnails/3.jpg)
What You Must Know:
The terminology of biotechnology.
The steps in gene cloning with special attention to the biotechnology tools that make cloning possible.
The key ideas that make PCR possible.
How gel electrophoresis can be used to separate DNA fragments or protein molecules.
![Page 4: Biotechnology Techniques How to make Recombinant DNA Gel Electrophoresis PCR Summarize: What is this technique? Draw and label a diagram to show this technique](https://reader030.vdocuments.us/reader030/viewer/2022032802/56649e0c5503460f94af5727/html5/thumbnails/4.jpg)
Genetic Engineering: process of manipulating genes and genomes
Biotechnology: process of manipulating organisms or their components for the purpose of making useful products.
![Page 5: Biotechnology Techniques How to make Recombinant DNA Gel Electrophoresis PCR Summarize: What is this technique? Draw and label a diagram to show this technique](https://reader030.vdocuments.us/reader030/viewer/2022032802/56649e0c5503460f94af5727/html5/thumbnails/5.jpg)
Recombinant DNA: DNA that has been artificially made, using DNA from different sources eg. Human gene inserted into E.coli
Gene cloning: process by which scientists can product multiple copies of specific segments of DNA that they can then work with in the lab
![Page 6: Biotechnology Techniques How to make Recombinant DNA Gel Electrophoresis PCR Summarize: What is this technique? Draw and label a diagram to show this technique](https://reader030.vdocuments.us/reader030/viewer/2022032802/56649e0c5503460f94af5727/html5/thumbnails/6.jpg)
Tools of Genetic Engineering
Restriction enzymes (restriction endonucleases): used to cut strands of DNA at specific locations (restriction sites) Restriction Fragments: have at least 1
sticky end (single-stranded end)
DNA ligase: joins DNA fragments
Cloning vector: carries the DNA sequence to be cloned (eg. bacterial plasmid)
![Page 7: Biotechnology Techniques How to make Recombinant DNA Gel Electrophoresis PCR Summarize: What is this technique? Draw and label a diagram to show this technique](https://reader030.vdocuments.us/reader030/viewer/2022032802/56649e0c5503460f94af5727/html5/thumbnails/7.jpg)
Using a restriction
enzyme (RE) and DNA ligase
to make recombinant
DNA
![Page 8: Biotechnology Techniques How to make Recombinant DNA Gel Electrophoresis PCR Summarize: What is this technique? Draw and label a diagram to show this technique](https://reader030.vdocuments.us/reader030/viewer/2022032802/56649e0c5503460f94af5727/html5/thumbnails/8.jpg)
Gene Cloning
![Page 9: Biotechnology Techniques How to make Recombinant DNA Gel Electrophoresis PCR Summarize: What is this technique? Draw and label a diagram to show this technique](https://reader030.vdocuments.us/reader030/viewer/2022032802/56649e0c5503460f94af5727/html5/thumbnails/9.jpg)
Applications of Gene Cloning
![Page 10: Biotechnology Techniques How to make Recombinant DNA Gel Electrophoresis PCR Summarize: What is this technique? Draw and label a diagram to show this technique](https://reader030.vdocuments.us/reader030/viewer/2022032802/56649e0c5503460f94af5727/html5/thumbnails/10.jpg)
Techniques of Genetic Engineering
![Page 11: Biotechnology Techniques How to make Recombinant DNA Gel Electrophoresis PCR Summarize: What is this technique? Draw and label a diagram to show this technique](https://reader030.vdocuments.us/reader030/viewer/2022032802/56649e0c5503460f94af5727/html5/thumbnails/11.jpg)
PCR (Polymerase Chain Reaction): amplify (copy) piece of DNA without use of cells
![Page 12: Biotechnology Techniques How to make Recombinant DNA Gel Electrophoresis PCR Summarize: What is this technique? Draw and label a diagram to show this technique](https://reader030.vdocuments.us/reader030/viewer/2022032802/56649e0c5503460f94af5727/html5/thumbnails/12.jpg)
Gel Electrophoresis: used to separate DNA molecules on basis of size and charge using an electrical current (DNA + pole)
![Page 13: Biotechnology Techniques How to make Recombinant DNA Gel Electrophoresis PCR Summarize: What is this technique? Draw and label a diagram to show this technique](https://reader030.vdocuments.us/reader030/viewer/2022032802/56649e0c5503460f94af5727/html5/thumbnails/13.jpg)
Gel Electrophoresis: used to separate DNA molecules on basis of size and charge using an electrical current (DNA + pole)
![Page 14: Biotechnology Techniques How to make Recombinant DNA Gel Electrophoresis PCR Summarize: What is this technique? Draw and label a diagram to show this technique](https://reader030.vdocuments.us/reader030/viewer/2022032802/56649e0c5503460f94af5727/html5/thumbnails/14.jpg)
Cloning OrganismsCloning Organisms
Nuclear transplantation: nucleus of egg is removed and replaced with nucleus of body cell
![Page 15: Biotechnology Techniques How to make Recombinant DNA Gel Electrophoresis PCR Summarize: What is this technique? Draw and label a diagram to show this technique](https://reader030.vdocuments.us/reader030/viewer/2022032802/56649e0c5503460f94af5727/html5/thumbnails/15.jpg)
Nuclear Transplantation
![Page 16: Biotechnology Techniques How to make Recombinant DNA Gel Electrophoresis PCR Summarize: What is this technique? Draw and label a diagram to show this technique](https://reader030.vdocuments.us/reader030/viewer/2022032802/56649e0c5503460f94af5727/html5/thumbnails/16.jpg)
Problems with Reproductive Cloning
Cloned embryos exhibited various defects
DNA of fully differentiated cell have epigenetic changes
![Page 17: Biotechnology Techniques How to make Recombinant DNA Gel Electrophoresis PCR Summarize: What is this technique? Draw and label a diagram to show this technique](https://reader030.vdocuments.us/reader030/viewer/2022032802/56649e0c5503460f94af5727/html5/thumbnails/17.jpg)
Stem Cells
Stem cells: can reproduce itself indefinitely and produce other specialized cells Zygote = totipotent (any type of cell) Embryonic stem cells = pluripotent (many cell
types) Adult stem cells = multipotent (a few cell types)
or induced pluripotent, iPS (forced to be pluripotent)
![Page 18: Biotechnology Techniques How to make Recombinant DNA Gel Electrophoresis PCR Summarize: What is this technique? Draw and label a diagram to show this technique](https://reader030.vdocuments.us/reader030/viewer/2022032802/56649e0c5503460f94af5727/html5/thumbnails/18.jpg)
Embryonic vs. Adult
stem cells
![Page 19: Biotechnology Techniques How to make Recombinant DNA Gel Electrophoresis PCR Summarize: What is this technique? Draw and label a diagram to show this technique](https://reader030.vdocuments.us/reader030/viewer/2022032802/56649e0c5503460f94af5727/html5/thumbnails/19.jpg)
Using stem cells for disease treatment
![Page 20: Biotechnology Techniques How to make Recombinant DNA Gel Electrophoresis PCR Summarize: What is this technique? Draw and label a diagram to show this technique](https://reader030.vdocuments.us/reader030/viewer/2022032802/56649e0c5503460f94af5727/html5/thumbnails/20.jpg)
Applications of DNA Technology
1. Diagnosis of disease – identify alleles, viral DNA
2. Gene therapy – alter afflicted genes
3. Production of pharmaceuticals
4. Forensic applications – DNA profiling
5. Environmental cleanup – use microorganisms
6. Agricultural applications - GMOs
![Page 21: Biotechnology Techniques How to make Recombinant DNA Gel Electrophoresis PCR Summarize: What is this technique? Draw and label a diagram to show this technique](https://reader030.vdocuments.us/reader030/viewer/2022032802/56649e0c5503460f94af5727/html5/thumbnails/21.jpg)
Gene therapy using a retroviral vector
![Page 22: Biotechnology Techniques How to make Recombinant DNA Gel Electrophoresis PCR Summarize: What is this technique? Draw and label a diagram to show this technique](https://reader030.vdocuments.us/reader030/viewer/2022032802/56649e0c5503460f94af5727/html5/thumbnails/22.jpg)
“Pharm” animal: produce human protein secreted in milk for medical
use
![Page 23: Biotechnology Techniques How to make Recombinant DNA Gel Electrophoresis PCR Summarize: What is this technique? Draw and label a diagram to show this technique](https://reader030.vdocuments.us/reader030/viewer/2022032802/56649e0c5503460f94af5727/html5/thumbnails/23.jpg)
DNA Fingerprinting
![Page 24: Biotechnology Techniques How to make Recombinant DNA Gel Electrophoresis PCR Summarize: What is this technique? Draw and label a diagram to show this technique](https://reader030.vdocuments.us/reader030/viewer/2022032802/56649e0c5503460f94af5727/html5/thumbnails/24.jpg)
RFLPs (“rif-lips”)
Restriction Fragment Length Polymorphism
Cut DNA with different restriction enzymes
Each person has different #s of DNA fragments created
Analyze DNA samples on a gel for disease diagnosis
Outdated method of DNA profiling (required a quarter-sized sample of blood)
![Page 25: Biotechnology Techniques How to make Recombinant DNA Gel Electrophoresis PCR Summarize: What is this technique? Draw and label a diagram to show this technique](https://reader030.vdocuments.us/reader030/viewer/2022032802/56649e0c5503460f94af5727/html5/thumbnails/25.jpg)
RFLPs – Disease Diagnosis
![Page 26: Biotechnology Techniques How to make Recombinant DNA Gel Electrophoresis PCR Summarize: What is this technique? Draw and label a diagram to show this technique](https://reader030.vdocuments.us/reader030/viewer/2022032802/56649e0c5503460f94af5727/html5/thumbnails/26.jpg)
STR Analysis
STR = Short Tandem Repeats
Non-coding DNA has regions with sequences (2-5 base length) that are repeated
Each person has different # of repeats at different locations (loci)
Current method of DNA fingerprinting used – only need 20 cells for analysis
![Page 27: Biotechnology Techniques How to make Recombinant DNA Gel Electrophoresis PCR Summarize: What is this technique? Draw and label a diagram to show this technique](https://reader030.vdocuments.us/reader030/viewer/2022032802/56649e0c5503460f94af5727/html5/thumbnails/27.jpg)
STR Analysis
![Page 28: Biotechnology Techniques How to make Recombinant DNA Gel Electrophoresis PCR Summarize: What is this technique? Draw and label a diagram to show this technique](https://reader030.vdocuments.us/reader030/viewer/2022032802/56649e0c5503460f94af5727/html5/thumbnails/28.jpg)
STR Analysis
![Page 29: Biotechnology Techniques How to make Recombinant DNA Gel Electrophoresis PCR Summarize: What is this technique? Draw and label a diagram to show this technique](https://reader030.vdocuments.us/reader030/viewer/2022032802/56649e0c5503460f94af5727/html5/thumbnails/29.jpg)