Ion Pair-Reversed Phase HPLC Approach Facilitates Subcloning of PCR Products and Screening of Recombinant Colonies | Zendy

Carla M. ShawBruha | Zendy; Kimberly A. Lamb | Zendy

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Ion Pair-Reversed Phase HPLC Approach Facilitates Subcloning of PCR Products and Screening of Recombinant Colonies

Author(s) -

Carla M. ShawBruha,

Kimberly A. Lamb

Publication year - 2000

Publication title -

biotechniques

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.617

H-Index - 131

eISSN - 1940-9818

pISSN - 0736-6205

DOI - 10.2144/00284pf02

Subject(s) - subcloning , recombinant dna , biology , cloning (programming) , high performance liquid chromatography , dna , molecular cloning , nucleic acid , microbiology and biotechnology , polymerase chain reaction , plasmid , chromatography , genetics , chemistry , gene , complementary dna , computer science , programming language

The isolation of a single DNA molecule for cloning is technically difficult, and the subsequent screening of colonies for recombinant DNA clones is time consuming. Ion pair-reversed phase HPLC (IP-RP-HPLC) analysis of nucleic acids improves the resolution and isolation of PCR products to be cloned. We demonstrate that PCR products analyzed and collected using the IP-RP-HPLC approach (WAVE DNA Fragment Analysis System) can be cloned directly into a plasmid vector. In addition, we demonstrate that when IP-RP-HPLC analysis is extended to the colony screening process, the time required for these procedures is reduced.

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