Methods in Biotechnology

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.  INDICE: 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 –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

• ISBN: 978-1-119-15678-9
• Editorial: Wiley–Blackwell
• Encuadernacion: Rústica
• Páginas: 360
• Fecha Publicación: 15/07/2016
• Nº Volúmenes: 1
• Idioma: Inglés