Abstract
This chapter deals with PCR analysis coupled to an ELISA-based detection system (PCR-ELISA). This type of assay ensures the specificity of the sequence of an amplified PCR product employing probes containing sequences complementary to an amplified target sequence. The method is advantageous to detect specific DNA and to differentiate among several PCR products. One of the possible applications is to discriminate between products amplified with one set of primers (consensus primers), but differing in the specific sequence spanned by these primer probes. In this chapter, a detailed description of the PCR-ELISA assay to detect Mycobacterium tuberculosis in formalin-fixed paraffin-embedded (FFPE) tissues is given.
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Notes
- 1.
EtBr is a potentially carcinogenic compound. Always wear gloves. Used EtBr solutions must be collected in containers for chemical waste and discharged according with the local hazardous chemical disposal procedures.
- 2.
Clean the pipette with alcohol or another disinfectant and leave them under the UV lamp for at least 10 min. Alternatively, it is possible to autoclave the pipette depending on the provider’s instructions for the different pipettes.
- 3.
Microtiter plates could also be used to run PCR.
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© 2011 Springer-Verlag Berlin Heidelberg
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Schewe, C., Weichert, W., Dietel, M. (2011). PCR-ELISA. In: Stanta, G. (eds) Guidelines for Molecular Analysis in Archive Tissues. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-17890-0_24
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DOI: https://doi.org/10.1007/978-3-642-17890-0_24
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