Taq DNA polymerase (with MgCl2 in Buffer) instruction manual
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1. Long-term storage (non-frequent use) at -70oC; stored at -20oC daily or weekly. Try to avoid multiple freeze-thaw cycles, do not expose to large temperature fluctuations, these fluctuations have a certain impact on product stability.
2. It is recommended that the enzyme be used in a concentration of 1 to 2.5 μl in 100 μl of the reaction solution. Excessive amounts of enzyme and excessive extension times may cause non-specific amplification bands.
3. To ensure the results of the experiment, it is recommended to use our company's supporting buffer and related reagents.
4. The amplification results are related to the reaction system, Mg2+ concentration, primers, and cycling parameters. In order to reduce unnecessary optimization process, save time and ensure stable and repeatable test results, we recommend using our PCR SmartMix.
Product Code: TQ-01/02/03/04
MyLab® Taq DNA Polymerase (with MgCl2 in Buffer)
Instruction manual "Specifications" 5U/ïl; 100ïl; 500U.
『Featuresã€
This preparation is a heat-resistant DNA polymerase having a molecular weight of 94 kDa. The gene of the Thermus aquaticus DNA Polymerase was cloned, expressed in Escherichia coli, and isolated and purified. It has the same function as the natural Taq DNA polymerase. The enzymatic reaction requires the participation of Mg2+, which uses double-stranded DNA as a template to catalyze the formation of double-stranded DNA from the 5' to the 3' end of the nucleotide. This enzyme does not have the activity of a 5'-3' exonuclease. This product is mainly used for PCR amplification of DNA, DNA sequencing and the like. The 3' end of the PCR product is A and can be used for T/A cloning. At 70-75oC, the rate of DNA polymerization is greater than 75 bases per second.
"composition"
Taq DNA Polymerase: 500U (5U/ïl; 200ïl)
10ï‚´ PCR Buffer: 1.5ml, the components are:
100 mM Tris-HCl (pH 8.7)
500 mM KCl
15 mM MgCl2
0.1% white gelatin other stabilizers and enhancers "active units"
One unit enzyme is defined as the amount of enzyme required to catalyze the addition of 10 nmol of an isotope-labeled dNTP to an acid-insoluble material at 72 ° C for 30 min.
"Storage conditions" are stored at -20 °C.
"Validity period" is 12 months.
"QC"
1. Overdigest (OD) detection: After 30 U of Taq DNA polymerase was reacted with 1 μg of λ DNA at 72 ° C for 16 hours, the degraded lambda DNA was not detected by agarose gel electrophoresis.
2. Notch enzyme assay: 30 U Taq DNA polymerase was reacted with supercoiled plasmid DNA at 72 ° C for 4 hours, and no nicking enzyme or linear DNA band was detected by agarose gel electrophoresis.
3. Thermal stability test: The half-life of each 2.5 U Taq DNA polymerase in buffer is longer than 1 hour at 92 °C.
4. The purity of SDS-PAGE is more than 99%.
Scope of application
Conventional PCR/reverse transcription PCR/fluorescence quantitative PCR/DNA sequencing/blend-end PCR product plus A and the like.
"Instructions"
Note: The following examples are for reference in most cases. The actual reaction conditions vary depending on the structure of the template, primers, etc., and the optimal reaction conditions need to be set according to actual conditions.
A 1 kb fragment was amplified using human genomic DNA as a template, and the reaction system was 25 μl.
10ï‚´PCR Buffer 2.5ïl
Primer 1 (10ïM) 1ïl
Primer 2 (10ïM) 1ïl
dNTP (10mM) 0.5ïl
Taq DNA Polymerase 1~2.5U
Template 1ïg
ddH2O is added to 25ïl
If the reaction system is different, the dosage can be increased or decreased according to this ratio. After mixing, place it in the PCR machine and run it according to the set program. After the reaction, 5 μl of the reaction product was taken and detected by agarose gel electrophoresis.
This product is for research use only. Do not use in medicine, clinical treatment, food, etc.