Ilya Taq DNA Polymerase Master Mix RED 2X
1.5 mM MgCl2 final concentration
– High performance thermostable Ilya Taq DNA polymerase
– Significantly reduced set up time
– Direct loading of products onto agarose gels
– Easy visualization of complete mixing
– Increased reproducibility
– Diminished risk of contamination
– No proofreading – lacks a 3′-5′ exonuclease activity
– Ideal for TA cloning – leaves an A overhang
Taq DNA Polymerase 2x Master Mix RED is a ready-to-use 2x reaction mix with the Ilya Taq DNA polymerase which is engineered by Ilya Biotech Co. Ltd. This master mix includes the NH4+ buffer system, dNTPs and magnesium chloride. Simply add primers, template and water to a total reaction volume of 50 μl to successfully carry out primer extensions and other molecular biology applications.
There is no need to buy and use separate loading dyes. Simply load a portion of the reaction product onto an agarose gel for electrophoresis and subsequent visualization.
Composition of the Taq DNA Polymerase 2x Master Mix RED (1.5 mM MgCl2 final concentration)
– Tris-HCl pH 8.5, (NH4)2S04, 3 mM MgCl2, 0.2% TweenÒ 20
– 0.4 mM of each dNTP
– 0.2 units/μl Ilya Taq DNA polymerase
– Red dye (inert)
Storage and Stability
Ilya Taq DNA Polymerase Master Mix RED 2X kit is stable at -20 °C for 2 years after the production date.
Taq DNA Polymerase is tested for contaminating activities, with no trace of endonuclease activity, nicking activity, exonuclease activity or priming activity.
One unit is defined as the amount of Ilya Taq DNA polymerase that incorporates 10 nmoles of dNTPs into acid-precipitable DNA in 30 minutes at 72 °C under standard assay conditions.
Suggested Protocol Using Ilya Taq Master Mix RED 2x
This protocol serves as a guideline for primer extensions. Optimal reaction conditions such as incubation times, temperatures, and amount of template DNA may vary and must be determined individually.
n The final MgCl2 concentration of this Taq Master Mix RED is 1.5 mM 2x. In some applications, more than 1.5 mM MgCl2 is required for best results. Use 25 mM MgCl2 (may be purchased separately) to adjust the Mg2+ concentration according to table1:
|Final MgCl2 concentration in reaction (mM)||1.5||2.0||2.5||3.0||3.5||4.0||4.5|
|Volume of 25 mM MgCl2||0||1||2||3||4||5||6|
- Thaw Ilya Taq Master Mix RED 2X. It is important to mix thoroughly. A Homogeneous solution should be used to avoid localized concentrations of salts.
- Prepare a reaction mix. Table 2 shows the reaction set up for a final volume of 50 μl.
|Ilya Taq Master Mix 2X||25 μl||1x|
|25 mM MgCl2||0 μl (0 – 7 μl)||1.5 mM (0.5 – 5 mM)|
|Primer A (10 μM)||1 μl (0.5 – 5 μl)||0.2 μM (0.1 – 1.0 μM)|
|Primer B (10 μM)||1 μl (0.5 – 5 μl)||0.2 μM (0.1 – 1.0 μM)|
|PCR-grade H2O||X μl||–|
|Template DNA||X μl||genomic DNA: 50 ng (10 – 500 ng)
plasmid DNA: 0.5 ng (0.1 – 1 ng)
bacterial DNA: 5 ng (1 – 10 ng)
|TOTAL volume||50 μl||–|