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.
By:
Seung-Beom Hong,
M. Bazlur Rashid,
Lory Z. Santiago-Vázquez
Imprint: Wiley-Blackwell
Country of Publication: United States
Dimensions:
Height: 277mm,
Width: 213mm,
Spine: 23mm
Weight: 1.021kg
ISBN: 9781119156789
ISBN 10: 1119156785
Pages: 368
Publication Date: 22 July 2016
Audience:
Professional and scholarly
,
Undergraduate
Format: Paperback
Publisher's Status: Active
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
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.
