Sequential Chemiluminescent Detection of Target DNAs without Stripping and Reprobing | Zendy

L. Vinod Kumar Reddy | Zendy; Renuka DeSilva | Zendy; R. Handley | Zendy; A. Paul Schaap | Zendy; Hashem Akhavan-Tafti | Zendy

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Sequential Chemiluminescent Detection of Target DNAs without Stripping and Reprobing

Author(s) -

L. Vinod Kumar Reddy,

Renuka DeSilva,

R. Handley,

A. Paul Schaap,

Hashem Akhavan-Tafti

Publication year - 1999

Publication title -

biotechniques

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.617

H-Index - 131

eISSN - 1940-9818

pISSN - 0736-6205

DOI - 10.2144/99264st08

Subject(s) - chemiluminescence , dna , microbiology and biotechnology , peroxidase , substrate (aquarium) , hybridization probe , southern blot , enzyme , blot , alkaline phosphatase , chemistry , biochemistry , biology , stripping (fiber) , chromatography , gene , materials science , ecology , composite material

We present a simple method for sequential chemiluminescent detections of two different DNA loci on a single Southern blot. First, an enzyme-linked DNA probe for a unique sequence is detected with a horse-radish peroxidase (HRP) substrate followed by the detection of another enzyme-linked DNA probe for a different unique sequence with an alkaline phosphatase (AP) substrate that simultaneously inhibits the chemiluminescence generated by HRP. Such sequential detection steps eliminate the need to strip and reprobe blots and can be performed with no intervening steps.

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