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The first edition of this book was the first manual for laboratory work in the rapidly expanding field of synthetic biology. Based upon a highly successful university course by one of the pioneers in synthetic biology, the manual became particularly popular with students of the enormous annual international Genetically Engineered Machine (iGEM) competition. Questions at the time included the scalability of BioBrick cloning, how to stabilize chromoprotein expression and change the colors, and how to adapt methods for high schools and biohackers. A decade later, this second edition answers these questions with huge BioBrick constructs (front cover), next-generation less-toxic chromoproteins in a kit, and ultraviolet-light-free quantitation by smartphones. Further updates include a computational modeling lab and new avenues in SynBio.