Methods in Biotechnology
Wiley-Blackwell (Verlag)
978-1-119-15678-9 (ISBN)
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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.
SEUNG-BEOM HONG is a Visiting Assistant Professor of Biotechnology at the University of Houston-Clear Lake in Houston, TX, USA. M. BAZLUR RASHID is an Associate Professor of Biology and Biotechnology and former Program Chair of Biotechnology at the University of Houston-Clear Lake in Houston, TX, USA. LORY Z. SANTIAGO-VÁZQUEZ is an Associate Professor of Biotechnology at the University of Houston-Clear Lake in Houston, TX, USA.
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 𝛃-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
| Erscheinungsdatum | 19.07.2016 |
|---|---|
| Verlagsort | Hoboken |
| Sprache | englisch |
| Maße | 213 x 277 mm |
| Gewicht | 1021 g |
| Themenwelt | Naturwissenschaften ► Biologie ► Mikrobiologie / Immunologie |
| Naturwissenschaften ► Chemie | |
| Technik ► Umwelttechnik / Biotechnologie | |
| ISBN-10 | 1-119-15678-5 / 1119156785 |
| ISBN-13 | 978-1-119-15678-9 / 9781119156789 |
| Zustand | Neuware |
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