Wet lab-course-lec2
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This document provides an overview of topics covered in a Wet-Lab course lecture on DNA, including the structure of DNA, DNA replication, plasmids, and polymerase chain reaction (PCR). It defines key terms like DNA, double helix, nucleotide, and discusses the basic process of DNA replication in prokaryotes. PCR is introduced as an in vitro method of DNA replication using heat-stable DNA polymerase, primers, and nucleotides to amplify a specific DNA sequence. The three main steps of PCR - denaturation, annealing of primers, and extension - are outlined.
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Wet lab-course-lec2
- 3. outline • DNA • Plasmid • DNA replication • PCR
- 5. DNA
- 8. DNA Double Helix Nucleotide Polar Hydrogen Bond Coding / Template Folate
- 9. DNA Double Helix Nucleotide Polar Hydrogen Bond Coding / Template Folate
- 10. DNA Double Helix Nucleotide Polar Hydrogen Bond Coding / Template Folate
- 11. DNA Double Helix Nucleotide Polar Hydrogen Bond Coding / Template Folate
- 12. DNA Nucleotide
- 15. DNA Double Helix Nucleotide Polar Hydrogen Bond Coding / Template Folate
- 16. Polar
- 17. Polar
- 18. DNA Double Helix Nucleotide Polar Hydrogen Bond Coding / Template Folate
- 19. DNA Double Helix Nucleotide Polar Hydrogen Bond Coding / Template Folate
- 20. DNA Coding / Template
- 21. DNA Double Helix Nucleotide Polar Hydrogen Bond Coding / Template Folate
- 22. Folate vit B9
- 23. DNA Double Helix Nucleotide Polar Hydrogen Bond Coding / Template Folate
- 24. Plasmid
- 25. Plasmid • Non-chromosome • Small • Circular • Double-stranded • DNA
- 26. Plasmid Transtission • Transformation • Transduction • Conjugation
- 27. Transformation
- 28. Transduction
- 29. Conjugation
- 30. DNA replication
- 32. 1. Identified the origin 2. Unwinding the DNA helix 3. Initiation 4. Elongation 5. Reconstitution of chromatin structure
- 33. (SSB)
- 35. Replication in Prokaryotes • Occurs in the cytoplasm • Only one origin in each prokaryotic DNA molecule one replication bubble • replication rate is very rapid
- 37. PCR the in vitro version of DNA Replication
- 38. • A heat-stable DNA Polymerase (e.g.Taq Polymerase) • Primers • dNTPs(Deoxynucleotide)
- 40. Step 1 Denaturation of DNA This occurs at 94~98 ºC mimicking the function of helicase in the cell.
- 41. Step 2 Annealing or Primers Binding Primers bind to the complimentary sequence on the target DNA. Primers are chosen such that one is complimentary to the one strand at one end of the target sequence and that the other is complimentary to the other strand at the other end of the target sequence. Forward Primer Reverse Primer
- 42. Step 3 Extension or Primer Extension DNA polymerase catalyzes the extension of the strand in the 5-3 direction, starting at the primers, attaching the appropriate nucleotide (A-T, C-G) extension extension
- 43. More about Primers • PCR primers are short, single stranded DNA molecules (15-40 bp) • Manufactured commercially • Sequence specific • Primers designed with a longer length (15 → 40 bp), the primers become more selective.
- 44. Types of PCR • Real time PCR (q-PCR) • Reverse Transcriptase PCR (RT-PCR) • Nested PCR