Abstract
When designing primers, it is of initial importance to define the target area, and secondly the type of application. The BLAST function from the National Center for Biotechnology Information (NCBI) will help to identify the most suitable gene sequence to be used. There are many software programs; some free on websites/pages on the internet, dedicated to primer design and primer optimisation. In this chapter the most important factors that need to be taken into consideration when designing and optimising primers are highlighted.
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Troubleshooting
http://www3.appliedbiosystems.com/cms/groups/mcb_support/documents/generaldocuments/cms_042997.pdf
http://www.protocol-online.org/prot/Molecular_Biology/PCR/Real-Time_PCR/
http://www.appliedbiosystems.com/support/tutorials/pdf/data_analysis_7700.pdf
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Pestana, E.A., Belak, S., Diallo, A., Crowther, J.R., Viljoen, G.J. (2009). Analysis and Troubleshooting. In: Early, rapid and sensitive veterinary molecular diagnostics - real time PCR applications. Springer, Dordrecht. https://doi.org/10.1007/978-90-481-3132-7_7
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DOI: https://doi.org/10.1007/978-90-481-3132-7_7
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