High-throughput, cost-effective verification of structural DNA assembly
| Autor: | Nicole Klinkner, Todd Slaby, Sunil S. Chandran, Elaine B. Shapland, Daniel Hollis, Kedar G. Patel, Yandi Dharmadi, Jed Dean |
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| Rok vydání: | 2013 |
| Předmět: |
Genetics
Cost effectiveness In silico Electrophoresis Capillary DNA DNA Restriction Enzymes Sequence Analysis DNA Computational biology Biology Polymerase cycling assembly Synthetic biology Plasmid Rolling circle replication Methods Online Computer Simulation Synthetic Biology Genomic library Restriction digest Algorithms |
| Zdroj: | Nucleic Acids Research |
| ISSN: | 1362-4962 0305-1048 |
| DOI: | 10.1093/nar/gkt1088 |
| Popis: | DNA ‘assembly’ from ‘building blocks’ remains a cornerstone in synthetic biology, whether it be for gene synthesis (∼1 kb), pathway engineering (∼10 kb) or synthetic genomes (>100 kb). Despite numerous advances in the techniques used for DNA assembly, verification of the assembly is still a necessity, which becomes cost-prohibitive and a logistical challenge with increasing scale. Here we describe for the first time a comprehensive, high-throughput solution for structural DNA assembly verification by restriction digest using exhaustive in silico enzyme screening, rolling circle amplification of plasmid DNA, capillary electrophoresis and automated digest pattern recognition. This low-cost and robust methodology has been successfully used to screen over 31 000 clones of DNA constructs at |
| Databáze: | OpenAIRE |
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