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Two step PCR method to exchange the resistance cassette of a vector

View ORCID ProfileNils Cremer, View ORCID ProfileAnne Diehl
doi: https://doi.org/10.1101/2021.01.28.428618
Nils Cremer
Department of NMR-supported structural Biology, Leibniz-Forschungsinstitut für Molekulare Pharmakologie (FMP), Robert-Rössle-Str. 10, 13125 Berlin, Germany
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Anne Diehl
Department of NMR-supported structural Biology, Leibniz-Forschungsinstitut für Molekulare Pharmakologie (FMP), Robert-Rössle-Str. 10, 13125 Berlin, Germany
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Abstract

For co-transformation of two plasmids, both have to possess different antibiotic selection markers. If that is not the case, normally the gene of interest (GOI) is subcloned into another vector. Here we introduce a fast and easy method to exchange the antibiotic resistance cassette (ARC) in only two PCR steps.

Method Summary To shuttle the antibiotic resistance cassette (ARC) from one vector to another, one can amplify the ARC of interest and use the resulting PCR-product as a primer pair for the next amplification step. Simply remove parental DNA template by DpnI digestion, transform PCR product directly in E. coli cells, select transformants on an appropriate agar plate and isolate target vector by plasmid preparation.

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted January 28, 2021.
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Two step PCR method to exchange the resistance cassette of a vector
Nils Cremer, Anne Diehl
bioRxiv 2021.01.28.428618; doi: https://doi.org/10.1101/2021.01.28.428618
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Two step PCR method to exchange the resistance cassette of a vector
Nils Cremer, Anne Diehl
bioRxiv 2021.01.28.428618; doi: https://doi.org/10.1101/2021.01.28.428618

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