Primer check
Priming site position
Sequencing primers are essential for all sequencing reactions. To obtain the best possible data, sequencing primers should bind to a position 50 to 100 bases 5' off the desired sequence start.
Storage
Sequencing primers as other material delivered by customers are stored at least for 1 month and can be used in following orders.
Calculating primer melting temperatures for DNA sequencing
The melting temperature calculation form available above requires JavaScript. A form free of Javascript is also available. Nucleotide bases can be entered applying the IUPAC base nomenclature. Up to 8 bases may be degenerated.
Efficient primer binding is facilitated by primer melting temperatures of 60°C to 65°C. For G+C rich templates the melting temperatures should be 62°C to 66°C. Primer melting temperatures depend on the salt and primer concentrations actually used in the sequencing reactions. Melting temperatures for IIT Biotech's sequencing protocols can be checked here.