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Rapid Verification of Terminators Using the pGR-Blue Plasmid and Golden Gate Assembly

JoVE Journal
Biology

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JoVE Journal Biology

Rapid Verification of Terminators Using the pGR-Blue Plasmid and Golden Gate Assembly

Rapid Verification of Terminators Using the pGR-Blue Plasmid and Golden Gate Assembly

DOI:

10.3791/54064-v

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09:51 min

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April 25, 2016

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Jace C. Bradshaw, Allea Belle Gongola, Nathan S. Reyna

¹Department of Biology,Ouachita Baptist University

Chapters

00:05Title
01:28Annealing Oligonucleotides (Applies Only to Freeze-dried DNA)
02:43Golden Gate Assembly (Cost Efficient Method Using a Standard Mix)
04:19Transformation and Colony Selection
05:19Verification and Quantification of Terminator
07:55Quantification of Six Different Terminators in pGR-Blue
09:02Conclusion

Summary

Automatic Translation

This protocol utilizes Golden Gate Assembly and the plasmid pGR-blue to rapidly quantify the strength of terminators found in silico.

Tags

Keywords: Terminator In-vitro Golden Gate Assembly Bio-informatics Rapid Verification PGR-Blue Plasmid Oligonucleotides Annealing Restriction Enzyme Ligase Cloning Efficiency

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