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What are the factors to consider when designing qPCR primers? 

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When designing qPCR primers, several factors need to be considered. These include primer sequence, length, and melting temperature (Tm) . Proper primer design is critical for the success of the qPCR assay, as primers are the main determinants of specificity, sensitivity, and robustness . The primer sequence should be carefully selected to ensure efficiency and specificity of the amplification reaction, resulting in a high yield of the desired amplicon . The length of the primers should be appropriate to facilitate efficient binding to the target sequence . The melting temperature (Tm) of the primers should be within a suitable range to ensure optimal annealing during the qPCR reaction . Additionally, it is important to consider potential issues such as primer dimer formation and competition with secondary structures at the binding sites . By taking these factors into account, the design of qPCR primers can be optimized for accurate and reliable amplification of the target sequence.

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Open accessJournal ArticleDOI
Stephen A. Bustin, Jim F. Huggett 
169 Citations
The paper discusses factors to consider when designing qPCR primers, such as specificity, formation of dimers, competition with template secondary structures, and hybridization within a narrow temperature range.
The provided paper is about designing primers for the amplification-refractory mutation system (ARMS-PCR) technique. It does not provide information about designing qPCR primers.
The provided paper does not specifically mention qPCR primers. The paper discusses the parameters for successful PCR primer design in general.
Book ChapterDOI
01 Jan 2022
The provided paper does not specifically mention qPCR primer design.
The provided paper does not specifically mention qPCR primers.

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