1. DNA-based technology
New and old technologies that are utilized in biotechnology
DNA cloning
DNA libraries
Polymerase chain reaction (PCR)
Genome sequencing
DNA microarray
DNA fingerprinting

2. DNA-based technology
DNA cloning - separating a specific gene or DNA segment, attaching it to a small molecule of carrier DNA, replicating lots of DNA
5 steps to cloning (recombinant) DNA technology
1. Cut DNA at precise locations
2. Select small molecule of DNA (cloning vector) that can self-replicate - cut
3. Join two DNA fragments covalently
4. Move recombinant DNA to host cells
5. Identify host cells with recombinant DNA

3. DNA-based technology

4. DNA-based technology

5. DNA-based technology
DNA libraries - collection of DNA clones, gathered together as a source of DNA for sequencing, gene discovery, or gene function studies
Different types of libraries available
genomic library
cDNA library
and more...

6. DNA-based technology Polymerase chain reaction (PCR) Denature dsDNA
Anneal primers to now ssDNA
Polymerization (DNA synthesis - get 2 copies from 1)
One cycle can be repeated over & over
After cycle 30, > 1 billion identical molecules (230 = 1.07 x 109)

7. DNA-based technology
Polymerase chain reaction (PCR)

8. DNA-based technology
Genome sequencing
Human Genome

9. DNA-based technology
DNA microarray

10. DNA Fingerprinting
Used for detection of genetic diseases, forensics, paternity, evolutionary links
Based on the characteristics of mammalian DNA
Eukaryotic genome
DNA divided into 3 classes
1. Nonrepetitive DNA - 50% of DNA, contains almost all the genes that code for proteins
2. Moderately repetitive DNA - contains a few genes, also ribosomal RNA, has long stretches of nongenic DNA
3. Highly repetitive DNA (satellite DNA) - no genes, variation in sequences between individuals is most pronounced in DNA that does not code for proteins
Hypervariable regions called “polymorphic sites”
SO .. Restriction fragment with a polymorphic site may differ from one person to the next resulting in: RESTRICTION FRAGMENT LENGTH POLYMORPHISMS (RFLPs)
Pattern of bands called a “DNA fingerprint” because can use it to identify an individual

11. DNA Fingerprinting

12. DNA Fingerprinting