Lambda‐PCR for precise DNA assembly and modification Article Swipe
Imen Tanniche
,
Amanda K. Fisher
,
Frank Gillam
,
Eva Collakova
,
Chenming Zhang
,
David R. Bevan
,
Ryan S. Senger
·
YOU?
·
· 2022
· Open Access
·
· DOI: https://doi.org/10.1002/bit.28240
YOU?
·
· 2022
· Open Access
·
· DOI: https://doi.org/10.1002/bit.28240
Lambda‐polymerase chain reaction (λ‐PCR) is a novel and open‐source method for DNA assembly and cloning projects. λ‐PCR uses overlap extension to ultimately assemble linear and circular DNA fragments, but it allows the single‐stranded DNA (ssDNA) primers of the PCR extension to first exist as double‐stranded DNA (dsDNA). Having dsDNA at this step is advantageous for the stability of large insertion products, to avoid inhibitory secondary structures during direct synthesis, and to reduce costs. Three variations of λ‐PCR were created to convert an initial dsDNA product into an ssDNA “megaprimer” to be used in overlap extension: (i) complete digestion by λ‐exonuclease, (ii) asymmetric PCR, and (iii) partial digestion by λ‐exonuclease. Four case studies are presented that demonstrate the use of λ‐PCR in simple gene cloning, simultaneous multipart assemblies, gene cloning not achievable with commercial kits, and the use of thermodynamic simulations to guide λ‐PCR assembly strategies. High DNA assembly and cloning efficiencies have been achieved with λ‐PCR for a fraction of the cost and time associated with conventional methods and some commercial kits.
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- Type
- article
- Language
- en
- Landing Page
- https://doi.org/10.1002/bit.28240
- https://onlinelibrary.wiley.com/doi/pdfdirect/10.1002/bit.28240
- OA Status
- hybrid
- Cited By
- 4
- References
- 39
- Related Works
- 10
- OpenAlex ID
- https://openalex.org/W4296808201
All OpenAlex metadata
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https://doi.org/10.1002/bit.28240Digital Object Identifier
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Lambda‐PCR for precise DNA assembly and modificationWork title
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articleOpenAlex work type
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enPrimary language
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2022Year of publication
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2022-09-23Full publication date if available
- Authors
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Imen Tanniche, Amanda K. Fisher, Frank Gillam, Eva Collakova, Chenming Zhang, David R. Bevan, Ryan S. SengerList of authors in order
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https://doi.org/10.1002/bit.28240Publisher landing page
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hybridOpen access status per OpenAlex
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Lambda, DNA, Computational biology, Biology, Genetics, Chemistry, Molecular biology, Physics, OpticsTop concepts (fields/topics) attached by OpenAlex
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10Other works algorithmically related by OpenAlex
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