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Full Description
As rapid advances in biotechnology occur, there is a need for a pedagogical tool to aid current students and laboratory professionals in biotechnological methods; Methods in Biotechnology is an invaluable resource for those students and professionals.
Methods in Biotechnology engages the reader by implementing an active learning approach, provided advanced study questions, as well as pre- and post-lab questions for each lab protocol. These self-directed study sections encourage the reader to not just perform experiments but to engage with the material on a higher level, utilizing critical thinking and troubleshooting skills.
This text is broken into three sections based on level - Methods in Biotechnology, Advanced Methods in Biotechnology I, and Advanced Methods in Biotechnology II. Each section contains 14-22 lab exercises, with instructor notes in appendices as well as an answer guide as a part of the book companion site. This text will be an excellent resource for both students and laboratory professionals in the biotechnology field.
Contents
Preface vii
Acknowledgements ix
Introduction to biotechnology lab xi
About the companion website xix
Part I: Methods in Biotechnology (MB) Laboratory Exercises 1
1 MB experiment 1: Lab measurements 3
2 MB experiment 2: Use of the spectrophotometer and Beer's law 9
3 MB experiment 3: Making solutions and buffer efficacy 15
4 MB experiment 4: Acid-base titration 19
5 MB experiment 5: Protein denaturation and precipitation 23
6 MB experiment 6: Bacterial transformation 27
7 MB experiment 7: GFP purification 33
8 MB experiment 8: SDS-PAGE analysis 37
9 MB experiment 9: DNA isolation 41
10 MB experiment 10: PCR-based Alu-human DNA typing 45
11 MB experiment 11: Restriction enzyme digestion 51
12 MB experiment 12: Agarose gel electrophoresis 53
13 MB experiment 13: Ouchterlony and ELISA immunoassays 57
14 MB experiment 14: Testing plant substances for antimicrobial activity 63
15 MB experiment 15: Peroxidase enzyme activity assay 67
Part II: Advanced Methods in Biotechnology (AMB) 1 Laboratory Exercises 71
16 AMB 1 experiment 16: Aseptic technique and culture handling 73
17 AMB 1 experiment 17: Yeast culture media preparation 77
18 AMB 1 experiment 18: Growth curve 79
19 AMB 1 experiment 19: Mini plasmid prep 83
20 AMB 1 experiment 20: Restriction digestion, purification concentration and quantification of DNA 87
21 AMB 1 experiment 21: Polymerase chain reaction (PCR) 91
22 AMB 1 experiment 22: TA blunt end SLIC, and CPEC cloning of PCR product 97
23 AMB 1 experiment 23: One-step multifragment assembly cloning 103
24 AMB 1 experiment 24: Restriction enzyme digestion and fast agarose gel electrophoresis 111
25 AMB 1 experiment 25: Southern blot transfer 117
26 AMB 1 experiment 26: Probe labeling and purification 121
27 AMB 1 experiment 27: Dot blot assay 125
28 AMB 1 experiment 28: Pre-hybridization, hybridization and detection 127
29 AMB 1 experiment 29: Total yeast RNA isolation and RT-PCR 131
30 AMB 1 experiment 30: Yeast-based in vivo recombination cloning 139
31 AMB 1 experiment 31: Plasmid DNA isolation from yeast 143
32 AMB 1 experiment 32: E. coli transformation with yeast plasmid DNA 145
33 AMB 1 experiment 33: X-gal filter lift assay 147
34 AMB 1 experiment 34: Protein quantitation assay 149
35 AMB 1 experiment 35: Quantitative 훃-Galactosidase assay in yeast 155
36 AMB 1 experiment 36: Gel filtration chromatography (GFC) 161
37 AMB 1 experiment 37: Ion exchange chromatography (IEC) 165
Part III: Advanced Methods in Biotechnology (AMB) 2 Laboratory Exercises 169
38 AMB 2 experiment 38: E. coli culture media preparation 171
39 AMB 2 experiment 39: Site-directed mutagenesis 175
40 AMB 2 experiment 40: Protein expression in E. coli 179
41 AMB 2 experiment 41: Protein purification by affinity column chromatography 187
42 AMB 2 experiment 42: SDS-PAGE analysis of affinity column fractions 193
43 AMB 2 experiment 43: Western blot analysis of affinity column fractions 199
44 AMB 2 experiment 44: Yeast media preparation and phenotypic analysis of yeast strains 203
45 AMB 2 experiment 45: Yeast transformation for yeast two-hybrid (Y2H) assay and genome editing by CRISPR-Cas system 209
46 AMB 2 experiment 46: Yeast mating-mediated Y2H assay and genomic PCR 215
47 AMB 2 experiment 47: Yeast colony PCR screening and cycle DNA sequencing 221
48 AMB 2 experiment 48: DNA sequencing electrophoresis 227
49 AMB 2 experiment 49: RNA interference 237
50 AMB 2 experiment 50: Protein preparation for 2D gel electrophoresis 241
51 AMB 2 experiment 51: Two-dimensional gel electrophoresis 245
Part IV: Appendices 253
1 Methods in Biotechnology Appendix 1 255
2 MB Appendix 2 261
3 MB Appendix 3 265
4 MB Appendix 4 267
5 AMB 1 Appendix 1 271
6 AMB 1 Appendix 2 281
7 AMB 1 Appendix 3 287
8 AMB 1 Appendix 4 291
9 AMB 1 Appendix 5 297
10 AMB 2 Appendix 1 299
11 AMB 2 Appendix 2 307
12 AMB 2 Appendix 3 313
13 AMB 2 Appendix 4 317
14 AMB 2 Appendix 5 319
15 AMB 2 Appendix 6 321
16 AMB 2 Appendix 7 325
Glossary 327
Abbreviations 335
Index 337
