Some applications of this product may require a license which is not provided by the purchase of this product.
For research use only.
HOT FIREPol® MultiPlex Mix Ready to Load is a premixed ready-to-use solution containing all reagents required for hot-start multiplex PCR (except template, primers, and water) and an additional compound needed for direct loading onto an agarose gel and two tracking dyes (blue and yellow) that allow monitoring progress during electrophoresis.
Generated PCR products are compatible with T/A cloning procedures.
We recommend using HOT FIREPol® MultiPlex Mix Ready To Load in any PCR application that will be visualized by agarose gel electrophoresis and ethidium bromide staining.
HOT FIREPol® MultiPlex Mix Ready To Load is not recommended for use in applications where spectrophotometric measurements (absorbance or fluorescence) are necessary because yellow and blue dyes can interfere with these applications.
Concentration: 5x
Hot-start: yes, initial activation in 12-15 min
Ready to load: yes
Fidelity: 1 x Taq
Cloning Type: T/A cloning
Multiplexing: tested with up to 18 targets (depending on amplicon length)
Amplicon Size: up to 5 kb
HOT FIREPol® DNA polymerase: chemically modified FIREPol® DNA Polymerase enabling hot-start
5x Multiplex Buffer with 10 mM MgCl2: 1x PCR solution – 2 mM MgCl2
dNTPs: dATP, dCTP, dGTP and dTTP
Bovine serum albumin (BSA) to enhance PCR reaction
Blue dye: migration equivalent to 3.5-4.5 kb DNA fragment
Yellow dye: migration equivalent to 35-45 bp DNA fragment
Compound that increases sample density for direct loading
Bovine serum albumine (BSA) is one of the PCR additives that is used to enhance PCR reaction.
It has been proved that BSA increases PCR yields from low purity templates containing iron chloride, hemin, fulminic acid, humic acid, tannic acid, stool extracts and melanin (Al-Soud and Rådström 2000; Scipioni et al. 2008b; Opel et al. 2010).
Moreover, BSA also improves specificity in amplification of regions with secondary structures. It is also reported to prevent reaction components from sticking to tube walls.
FIREPol® and HOT FIREPol® DNA Polymerases and Master Mixes, as well as SolisFAST® Master Mix are compatible with most restriction enzymes without cleaning up the PCR reaction product.
SolisFAST® Master Mix with UNG contains dUTP instead of dTTP leading to the generation of PCR products containing uracil. Check your restriction enzyme for the ability to digest U-containing DNA amplicons.
Products generated by our FIREPol®, HOT FIREPol® DNA Polymerases and Master Mixes, as well as SolisFast® Master Mixes may be used for Sanger sequencing.
Prior to the sequencing, the products should be purified to remove excess primers and nucleotides (enzymatically with ExoI/SAP or Spin column-based nucleic acid purification).
Products generated with 5x HOT FIREPol® Blend Master Mix should be cleaned up utilising column-based nucleic acid purification method, because this mix contains proofreading enzyme with 3’-5’ exonuclease activity and may degrade your sequencing primer.
Please note that Ready to Load versions of our Master Mixes are not recommended for use in applications where spectro-photometric measurements (absorbance or fluorescence) are necessary because tracking dyes can interfere with these applications.
Yes, Solis BioDyne HOT FIREPol® DNA Polymerase and 5x HOT FIREPol® Blend Master Mix are suitable for colony PCR to determine the presence or absence of insert DNA in plasmid constructs directly from bacterial colonies without culturing or plasmid purification steps.
Sometimes, poor quality of the colony material (e.g. presence of contaminants) could inhibit the PCR reaction. It is important to use freshly grown colonies (overnight growth). If possible, include a positive (e.g. purified plasmid DNA with a desired insert) and a negative control (containing all components except template DNA) in your experimental set-up.
Recommended final amount of cDNA sample in downstream PCR reaction is up to one tenth of the final reaction volume. Overload of cDNA sample may compromise the downstream PCR, because cDNA sample may contain reaction components that may inhibit your PCR reaction.
Solis BioDyne products should be stored at -20°C.
Shipping and temporary storage for up to 1 month at room temperature (*15−25°C) has no detrimental effects on the quality of Solis BioDyne reagents.
Freeze-thaw stability is tested for each product. Most PCR and qPCR products have passed 30 freeze-thaw cycles with no changes in performance. Specific information is found in Storage and Shipping conditions of each product.
When stored and handled under the recommended conditions, full activity of the reagents is retained until the Expiry Date printed on the tube label.
*World Health Organization (2003). Guidelines for the Storage of Essential Medicines and Other Health Commodities.
Routine storage: -20ºCTemporary storage for up to 1 month at room temperature has no detrimental effects on the quality of HOT FIREPol® Multiplex Mix Ready to Load
At room temperature
Hot-start DNA Polymerase with unique 30-day room temperature stability for your everyday PCR needs.
High performance hot-start Master Mix for efficient multiplex reaction - analyse up to 18 targets in one reaction.
Robust and reliable hot-start Master Mix with higher fidelity and longer amplification range for more demanding reactions. Includes two tracking dyes.
Convenient ready-to-load molecular weight marker to determine the size of dsDNA during gel electrophoresis. Contains 13 discrete bands ranging from 250 bp to 10 000 bp.
Convenient ready-to-load molecular weight marker to determine the size of dsDNA during gel electrophoresis. Contains 13 discrete bands ranging from 100 bp to 3000 bp.
