BioThermStarMix™ is a 2.5x concentrated reagent mix for Hot Start PCR comprising BioThermStar™ DNA Polymerase (0.06 u/µl), 2.5x PCR-Buffer with 3.75 mM MgCl2, 500µM of each dNTP and stbilizers. The 2.5x concentration of the Mix provides high versatility of setting-up Hot Start PCR reactions. Up to 60% of the final reaction volume can be used for the addition of primer and template DNA solutions, co-solvents or other additives, if necessary.
BioThermStarMix™ is suitable and tested for amplification of genomic targets ranging from 100 bp to 4 kb and of episomal targets (lambda phage;plasmids) up to 10 kb under various reaction conditions.
- high through-put Hot Start PCR
-routine diagnostic Hot Start PCR requiring high reproducibility
- DNA sequencing template preparation
BioThermStarMix™ can be stored at either -20°C in a freezer or at 2-8°C in ausual refrigerator. Shipment at ambient temperatures is possible without reduction of Hot Start PCR performance and activity. Storage at 2-8°C is convenient for easy and time-saving assembling of the Hot Start PCR assays. Storage of BioThermStarMix™ at -20°C is recommended for long-term storage after the mix has been used once under non-sterile conditions. Multiple freezing and thawing do not affect the performance or activity of the mix. At 2-8°C the BioThermStarMix™ is at least stable for 12 month, in frozen state for 2 years.
Do not contaminate the BioThermStarMix™ with primers and template DNA used in individual reactions.
Thaw and mix all components thoroughly, spin down shortly and chill on ice.
It is very important to mix the BioThermStarMix™ before use to avoid localized concentration!
1. Place the Hot Start tubes on ice. 2.Prepare first a template/primer mix according to the volumes given in the table below for different reaction volumes. Mix the template/primer mix and chill on ice.
3. Dispense noe the corresponding volume of BioThermStarMix™ (20 µl for a 50 µl reaction) followed by the template/primer mix into each reaction tube. Close tubes and mix well. Spin down shortly, if necessary, and chill the tubes on ice.
4. Start the Hot Start program. Transfer the tubes directly from the ice into the termal cycler when the temperature of the block has reached 95°C. Incubate ca. 7 min. at 95°C before cycling.
The stabilizers in the BioThermStarMix™ are potential growth substrates of bacterial contaminations! If the Mix has been opened and used under non-sterile conditions, the residual moiety should be used during the next 2 weeks with intermediate storage at 2-8°C or should be frozen at -20°C for longer storage!