1 Cell Biology Experiment 1

1 Understanding Cell Biology Experiments 2
2 Experiments using prokaryotic cells 3
Experiment 1: Preparation and sterilization of microbial growth medium 4
Experiment 2 Bacterial growth curve 9
Experiment 3: Antibiotic Susceptibility Testing Using DISC 14
Experiment 4: Production of competent cells using E. coli 18
Experiment 5: Heat-shock transformation of E. coli 22
Experiment 6 Isolation of Plasmid DNA (mini-preparation of plasmid DNA) 26
3 Experiments Using Eukaryotic Cells 30
Experiment 7: Culturing Animal Cells 31
Experiment 8 Cell Viability Measurement Experiment (MTT Assay) 37
Experiment 9: Observation of DNA Laddering Due to Apoptosis Induction 41
RNA extraction and cDNA synthesis from cell lines 10 and 45
Experiment 11 Cell isolation from rat spleen tissue 50
Experiment 12: Extraction of genomic DNA from animal tissues 54
4 The Significance and Utilization of Cell Biology Experiments 58

2 Biochemistry Experiment 59

1 Understanding Biochemistry Experiments 60
Experiment 1: Overview of Biomolecule Extraction and Analysis Methods 60
2 Protein extraction and quantification from animal cells 65
Experiment 2 Whole cell extraction 65
Experiment 3: Extraction of cytoplasmic and nuclear proteins using a storage-grade cell lysis buffer 69
Experiment 4: Measuring Protein Concentration Using a Spectrophotometer and Bradford Reagent 73
3 Analysis of protein expression using Western blotting 78
Experiment 5: Preparation of polyacrylamide gel 78
Experiment 6 Protein Electrophoresis and Gel Staining 84
Experiment 7 Western Blotting and Ponshu Staining 88
Experiment 8: Film Development and Protein Expression Analysis 94
4 Observation of polymorphism in cancer cell nuclei and DNA using a fluorescence microscope 98
Experiment 9: Using a Fluorescence Microscope 98
Experiment 10: Observation of Polymorphism in Cancer Cell Nuclei and DNA Using a Fluorescence Microscope 104
5 Genomic DNA Extraction and Analysis of DNA Damage Recovery Rates Induced by UV Light 108
Experiment 11: Induction of DNA Damage by UV Light 108
Experiment 12: Isolation of Genomic DNA 111
Experiment 13: Measuring DNA Concentration Using Nanodrops 115
Experiment 14: Measuring DNA Damage Repair Rates 118
6 The Significance and Utilization of Biochemical Experiments 123

3 Genomics Experiment 125

1 Understanding Genomics Experiments 126
Experiment 1 Orientation 128
Experiment 2: Calculating Reagent Concentrations 131
2 DNA typing 133
Experiment 3 Isolation of genomic DNA from peripheral blood 133
Experiment 4: Application of DNA Typing Using PCR Amplification 138
Experiment 5: Determining heterozygosity in the VNTR region 145
3 TA cloning technology 150
Experiment 6 PCR amplification of specific regions to be used for TA cloning 150
Experiment 7: Confirmation of PCR product for TA and purification of PCR product 154
Experiment 8 TA-ligation and bacterial transformation 157
Experiment 9 DNA isolation and restriction enzyme treatment from TA-cloned transformants 163
4 Recombinant DNA technology 171
Experiment 10 PCR of cDNA for Expression Vector 171
Experiment 11 Restriction Enzyme Treatment of Vector and Insert DNA 181
Experiment 12 Gel extraction and ligation reaction 186
Experiment 13: Ligation Verification and Transformation 190
Experiment 14: Transformant colony identification and colony PCR 193
Experiment 15 Recombinant DNA Isolation and Restriction Enzyme Treatment 197
Experiment 16 Recombinant DNA sequencing analysis 204
Search 209