Successful completion of a semi-automated enzyme-free cloning method
| Autor: | Domenico Maione, Stefano Bonacci, Scilla Buccato, Roberto Petracca |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Computer science Genetic Vectors Genomics Enzyme free Successful completion Molecular cloning Bioinformatics Polymerase Chain Reaction 01 natural sciences Biochemistry Set (abstract data type) 03 medical and health sciences Structural Biology Genetics Cloning Molecular Automation Laboratory Cloning business.industry 010401 analytical chemistry General Medicine Automation 0104 chemical sciences 030104 developmental biology Embedded system Laboratory automation business |
| Zdroj: | Journal of Structural and Functional Genomics. 17:57-66 |
| ISSN: | 1570-0267 1345-711X |
| DOI: | 10.1007/s10969-016-9207-z |
| Popis: | Nowadays, in scientific fields such as Structural Biology or Vaccinology, there is an increasing need of fast, effective and reproducible gene cloning and expression processes. Consequently, the implementation of robotic platforms enabling the automation of protocols is becoming a pressing demand. The main goal of our study was to set up a robotic platform devoted to the high-throughput automation of the polymerase incomplete primer extension cloning method, and to evaluate its efficiency compared to that achieved manually, by selecting a set of bacterial genes that were processed either in the automated platform (330) or manually (94). Here we show that we successfully set up a platform able to complete, with high efficiency, a wide range of molecular biology and biochemical steps. 329 gene targets (99 %) were effectively amplified using the automated procedure and 286 (87 %) of these PCR products were successfully cloned in expression vectors, with cloning success rates being higher for the automated protocols respect to the manual procedure (93.6 and 74.5 %, respectively). |
| Databáze: | OpenAIRE |
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