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The default value is 50.0 and the value may range from 0.1 to 50.0. This value is used in the calculation of primer melting temperature. Primer concentration specifies the nM concentration of primer DNA in the reaction.
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This value is used in the calculation of both primer and PCR product melting temperatures. Reaction Conditions Salt concentration specifies the mM salt concentration in the reaction.
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PCR product melting temperatures are calculated by using the formula of Baldino, et al. The maximum product length to enter is 5,000 bases. Primer melting temperatures is calculated by using the nearest-neighbor model of Borer, and thermodynamic parameters for DNA nearest-neighbor interactions and the salt dependence of oligonucleotides determined by SantaLucia Amplicon Criterias For PCR products, you can specify a range of acceptable product sizes and define the minum and maximum GC content and melting temperature (Tm). The maximum primer length you can search for are 100 bases. Primer Criterias For the PCR primer pairs you can specify minimum and maximum primer length, primer GC content and primer melting temperature (Tm). For PCR primer pairs, you can specify any required bases at the 3' end of the primer (3' clamp), and a maximum difference in primer melting temperatures. Design Parameters You can design PCR primers from the whole template (= target sequence) or limit the choices to a particular region. The template sequence may contain ambiguous bases, but the design tool will not select primers complementary to any ambiguous sites on the template sequence. Line breaks and blank spaces are allowed. DNA sequence information as well as FASTA sequences ( starting with an ">" followed by the name) are possible. Target Sequence Copy & paste the target sequence from an external source. You either can use the default constraint values or modify those values to customise the analysis. In selecting appropriate primers, a variety of constraints on the primer and amplified product sequences are already considered and taken as default values. The PCR primer desgin tool analyses the entered DNA sequence and chooses the optimum PCR primer pairs. Eurofins Genomics' PCR Primer Design Tool is using Prime+ of the GCG Wisconsin Package originally written by Irv Edelman.