DNA-based technologyNew and old technologies that are utilized in biotechnology

DNA cloningDNA librariesPolymerase chain reaction (PCR)Genome sequencingDNA microarrayDNA fingerprinting

DNA-based technologyDNA cloning - separating a specific gene or DNA segment, attaching it to asmall molecule of carrier DNA, replicating lots of DNA

5 steps to cloning (recombinant) DNA technology

1. Cut DNA at precise locations2. Select small molecule of DNA (cloning vector) that can self-replicate - cut3. Join two DNA fragments covalently4. Move recombinant DNA to host cells5. Identify host cells with recombinant DNA

DNA-based technology

DNA-based technology

DNA-based technology

DNA libraries - collection of DNA clones, gathered together as a source ofDNA for sequencing, gene discovery, or gene function studies

Different types of libraries availablegenomic librarycDNA libraryand more...

DNA-based technologyPolymerase chain reaction (PCR)Denature dsDNAAnneal primers to now ssDNAPolymerization (DNA synthesis - get 2 copies from 1)

One cycle can be repeated over & overAfter cycle 30, > 1 billion identical molecules (230 = 1.07 x 109)

DNA-based technology

Polymerase chain reaction (PCR)

DNA-based technology

Genome sequencing

Human Genome

DNA-based technologyDNA microarray

Used for detection of genetic diseases, forensics, paternity, evolutionary linksBased on the characteristics of mammalian DNA

Eukaryotic genomeDNA divided into 3 classes1. Nonrepetitive DNA - 50% of DNA, contains almost all the genes that code forproteins

2. Moderately repetitive DNA - contains a few genes, also ribosomal RNA, has longstretches of nongenic DNA

3. Highly repetitive DNA (satellite DNA) - no genes, variation in sequences betweenindividuals is most pronounced in DNA that does not code for proteinsHypervariable regions called “polymorphic sites”

SO .. Restriction fragment with a polymorphic site may differ from one person to thenext resulting in: RESTRICTION FRAGMENT LENGTH POLYMORPHISMS(RFLPs)Pattern of bands called a “DNA fingerprint” because can use it to identify an individual

DNA Fingerprinting

DNA Fingerprinting

DNA Fingerprinting