#Snapgene overlapping primers software
#Snapgene overlapping primers free

Front Environ Sci Eng 5:28–39Ĭhuang LY, Cheng YH, Yang CH (2013) Specific primer design for the polymerase chain reaction. Lim J, Shin SG, Lee S et al (2011) Design and use of group-specific primers and probes for real-time quantitative PCR. Raymaekers M, Smets R, Maes B et al (2009) Checklist for optimization and validation of real-time PCR assays. Yu Y, Lee C, Kim J et al (2005) Group-specific primer and probe sets to detect methanogenic communities using quantitative real-time polymerase chain reaction. Qiagen (2010) Critical factors for successful real-time PCR. Nolan T, Hands RE, Bustin SA (2006) Quantification of mRNA using real-time RT-PCR.

#Snapgene overlapping primers free

Thornton B, Basu C (2011) Real-time PCR (qPCR) primer design using free online software. Heid CA, Stevens J, Livak KJ et al (1996) Real time quantitative PCR. Holland PM, Abramson RD, Watson R et al (1991) Detection of specific polymerase chain reaction product by utilizing the 50–30 exonuclease activity of Thermus aquaticus DNA polymerase. Rosadas C, Cabral-Castro MJ, Vicente AC et al (2013) Validation of a quantitative real-time PCR assay for HTLV-1 proviral load in peripheral blood mononuclear cells. Rodríguez-Lázaro D, Hernández M (2013) Real time PCR in food science: introduction. Invitrogen (2008) Real-time PCR: from theory to practice. This chapter provides guidelines for the design of suitable primers and probes and their subsequent validation through the development of singlex qPCR, multiplex qPCR, and RT-qPCR protocols. Specific consideration should be taken into account when designing primers and probes for multiplex qPCR and reverse transcription qPCR (RT-qPCR). After a proper design, a precise validation of the primers and probes is necessary.

These tools often consider the default requirements for primers and probes, although new research advances in primer and probe design should be progressively added to different algorithm programs.

#Snapgene overlapping primers software

To design primers and probes for qPCR, multiple software programs and websites are available being numerous of them free. This factor is especially important when designing primers for SYBR ® Green protocols but also in designing probes to ensure specificity of the developed qPCR protocol.

The formation of primer-dimers and other non-specific products should be avoided or reduced. Design of primers and probes should meet several criteria to find potential primers and probes for specific qPCR assays. Design of primers and probes is one of the most crucial factors affecting the success and quality of quantitative real-time PCR (qPCR) analyses, since an accurate and reliable quantification depends on using efficient primers and probes.