BioTherm™ Mix is a 2.5x concentrated reagent mix for PCR comprising BioTherm™ DNA polymerase(0.06 u/µl), 2.5x PCR-Buffer with 3.75 mM MgCl2 , 500 µM of each dNTP and stabilizers. The 2.5x concentration of the Mix provides high versatility for setting-up 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.
BioTherm™ Mix is suitable and tested for amplification of genomic targets ranging from l00 bp to 4 kb and of episomal targets (lambda phage; plasmids) up to 10 kb under various reaction conditions.
- high through-put PCR
- routine diagnostic PCR requiring high reproducibility
- DNA sequencing template preparation
BioTherm™ Mix can be stored at either -20°C in a freezer or at 2-8°C in a usual refrigerator. Shipment at ambient temperature is possible without reduction of PCR performance and activity. Storage at 2-8°C is convenient for easy and time-saving assembling of the PCR assays. Storage of BioTherm™ Mix at -20°C is recommended for long-term storage after the mix has been sed once under non-sterile conditions. Multiple free-zing and thawing do not affect the performance or activity of the Mix.
At 2-8°C the BioTherm™ Mix is at least stable for 12 months, in frozen state for 2 years.
The stabilizers in the BioTherm™ Mix are potential growth substrates for 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!
Do not contaminate the BioTherm™ Mix 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 BioTherm™ Mix before use to avoid localized concentration!
- Place the PCR tubes on ice.
- 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.
- Dispense now the corresponding volume of BioTherm™ Mix (20 µl for a 50
µl reaction) followed by the tem-plate/primer mix into each reaction tube.
Close tubes and mix well. Spin down shortly, if necessary, and chill the tubes
- Start the program on the thermal cycler. Transfer the tubes directly from
ice into the thermal cycler when the temperature of the block has reached