Anthony C Forster,Erik Gullberg,Josefine Liljeruhm

Synthetic Biology

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Synthetic Biology: A Lab Manual is the first manual for laboratory work in the new and rapidly expanding field of synthetic biology. Aimed at non-specialists, it details protocols central to synthetic biology in both education and research. In addition, it provides all the information that teachers and students from high schools and tertiary institutions need for a colorful lab course in bacterial synthetic biology using chromoproteins and designer antisense RNAs. As a bonus, practical material is provided for students of the annual international Genetically Engineered Machine (iGEM) competition. The manual is based upon a highly successful course at Sweden's Uppsala University and is coauthored by one of the pioneers of synthetic biology and two bioengineering postgraduate students.
An inspiring foreword is written by another pioneer in the field, Harvard's George Church:“Synthetic biology is to early recombinant DNA as a genome is to a gene. Is there anything that SynBio will not impact? There was no doubt that the field of SynBio needed ‘A Lab Manual’ such as the one that you now hold in your hands.”
Contents:Introduction:What is Synthetic Biology, Exactly?The iGEM OutbreakA Synthetic Biology Lab ManualGenes, Chromoproteins and Antisense RNAs:E. coli DNA: Chromosomes, Plasmids and Copy NumberCoupling of Transcription and Translation in BacteriaPromoter and Terminator for TranscriptionRibosome Binding Site (RBS)Codon BiasChromoproteinsSmall Regulatory RNAs (sRNAs)Lab Rooms and Equipment:The Physical Lab SpacesEquipmentSafety is Priority #1:FiresChemicalsBiological Safety and DisposalDangerous EquipmentLab Course Projects:Time and ResourcesProject Overview and Learning ObjectivesThe Lab NotebookLab Section 1. Preparation of Chemical Solutions and Agar PlatesLab Section 2. Coloring Bacteria by Adding a Promoter to a Chromoprotein GeneLab Section 3: Rational Engineering of Chromoprotein Expression LevelLab Section 4. Other ExperimentsThe “Dreaded” ExamProtocols:IntroductionProtocol 1. Preparation of Solutions and Agar PlatesProtocol 2. Overnight Cultures with Antibiotics, and Glycerol StocksProtocol 3. BioBrick™ 3A Assembly and Gel AnalysisProtocol 4. Agarose Gel ElectrophoresisProtocol 5. Preparation of Competent E. coli Cells Using CaCl2Protocol 6. Transformation of CaCl2-Competent E. coli CellsProtocol 7. Bacterial Re-Streaking TechniquesProtocol 8. Lysis of E. coli Cells with LysozymeProtocol 9. Polymerase Chain Reaction (PCR)Protocol 10. Inverse PCR MutagenesisProtocol 11. Colony PCRProtocol 12. Gibson AssemblyAdvanced Methods:Flow Cytometry and Cell SortingRecombination in Plasmids and the ChromosomeElectrocompetent CellsThe International Genetically Engineered Machine (iGEM) Competition:How to Start an iGEM TeamUppsala iGEM 2011 — Show Color with ColorUppsala iGEM 2012 — Resistance is FutileUppsala iGEM 2013 — Lactonutritious — It's DeliciousAppendicesReadership: Students and researchers in biotechnology, cell/molecular biology and genetics.
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184 printed pages
Original publication
2014

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