Tools and Techniques of Molecular Biology

Molecular Biology · Basic Sciences

How it begins

Análisis exhaustivo y técnicamente riguroso de los métodos, fundamentos moleculares, protocolos, variantes y aplicaciones clínico-diagnósticas de las principales herramientas de la biología molecular moderna.

What it covers

1. Tools and Techniques of Molecular Biology
2. Introduction and Historical Context
3. Properties of Nucleic Acids Relevant to Molecular Biology Techniques
4. · DNA structure and complementarity
5. · Melting temperature (Tm)
6. Extraction of DNA, RNA and Proteins
7. · Genomic DNA extraction
8. · Total RNA extraction
9. · Protein extraction
10. Quantification and Quality Control of Nucleic Acids
11. · UV Spectrophotometry (NanoDrop)
12. · Fluorimetry (Qubit)
13. · Quality control electrophoresis
14. Polymerase Chain Reaction (PCR): Fundamentals and Variants
15. · Principle of PCR
16. · Phases of the PCR cycle
17. · PCR variants
18. Quantitative PCR (qPCR / RT-qPCR) and Digital PCR (ddPCR)
19. · Quantitative real-time PCR (qPCR)
20. · Digital PCR (dPCR / ddPCR)
21. Gel Electrophoresis — DNA and RNA
22. · Fundamentals of electrophoresis
23. · RNA electrophoresis
24. SDS-PAGE and Protein Electrophoresis
25. · SDS-PAGE (Sodium Dodecyl Sulfate–Polyacrylamide Gel Electrophoresis)
26. Southern, Northern and Western Blot
27. · Western Blot — Detailed protocol
28. FISH and In Situ Hybridization
29. · Fluorescent in situ hybridization (FISH)
30. · M-FISH and SKY: spectral karyotype
31. Sanger Sequencing (String Termination Sequencing)
32. · Rationale
33. Next Generation Sequencing (NGS / Massively Parallel Sequencing)
34. · Concept and revolution of NGS
35. · General principle of NGS (Illumina platform – sequencing by synthesis)
36. · Types of NGS experiments
37. Third Generation Sequencing — Long Reads
38. · PacBio (SMRT Sequencing)
39. · Oxford Nanopore (Nanopore Sequencing)
40. Restriction Enzymes
41. · Basis and classification
42. Molecular Cloning
43. · Molecular cloning concept
44. · Modern ligationless cloning strategies
45. Expression Vectors and Expression Systems
46. · Viral vectors for gene transfer
47. Transformation and Transfection
48. CRISPR-Cas9 and Variants — Precision Genome Editing
49. · Fundamentals of the CRISPR-Cas9 system
50. · DSB repair mechanisms and editing results
51. · Advanced variants of the CRISPR system
52. Zinc Finger Nucleases (ZFN) and TALEN
53. DNA and RNA microarrays
54. · Fundamentals of microarrays
55. RNA-seq and Transcriptomics
56. · RNA-seq workflow
57. · Single-cell RNA-seq (scRNA-seq)
58. Proteomics and Mass Spectrometry
59. · Mass spectrometry (MS) for proteomics
60. ChIP-seq, Epigenomics and ATAC-seq
61. · ChIP-seq (Chromatin Immunoprecipitation Sequencing)
62. · DNA methylation — sodium bisulfite
63. Immunofluorescence, IHC and Advanced Microscopy
64. · Immunohistochemistry (IHC) and immunofluorescence (IF)
65. · Advanced microscopy
66. Study of Molecular Interactions: Double Hybrid, FRET, Co-IP and SPR
67. Diagnostic Applications of Molecular Biology
68. · Molecular diagnosis of infectious diseases
69. · Molecular diagnosis in oncology
70. Gene Therapy, Interfering RNA (RNAi) and Therapeutic mRNA
71. · RNA interference (RNAi)
72. · Antisense oligonucleotides (ASO)