• Lyophilized Fast DNA TaqMan PCR Kit(UNG)
  • Lyophilized Fast DNA TaqMan PCR Kit(UNG)
  • Lyophilized Fast DNA TaqMan PCR Kit(UNG)
  • Lyophilized Fast DNA TaqMan PCR Kit(UNG)

Lyophilized Fast DNA TaqMan PCR Kit(UNG)

The TaqMan PCR Reagent in this kit is a freeze-dried reagent that uses hot start Fast Taq DNA polymerase, dNTP, etc. to make freeze-dried reagents. The product has stable performance and can be stored for a long time. When adding 2 × After the TaqMan Buffer is dissolved, the reagent becomes a conventional TaqMan PCR Mix single component premixed reagent. When conducting experiments, the preparation of the PCR reaction solution is very convenient and simple, simply adding primers, probes, DNA
$3.00
  • Lyophilized Fast DNA TaqMan PCR Kit(UNG)
  • Lyophilized Fast DNA TaqMan PCR Kit(UNG)

SPECIFICATION

This reagent kit is a specialized reagent for real-time fluorescence quantitative PCR using the TaqMan probe method. It is suitable for dual labeled probes such as FAM/HEX/TT/JOE/ROX (this product is also suitable for Molecular Beacon probes), and is widely used for relative quantitative detection of DNA and cDNA, gene detection, and development of gene diagnostic reagents.
The TaqMan PCR Reagent in this kit is a freeze-dried reagent that uses hot start Fast Taq DNA polymerase, dNTP, etc. to make freeze-dried reagents. The product has stable performance and can be stored for a long time. When adding 2 × After the TaqMan Buffer is dissolved, the reagent becomes a conventional TaqMan PCR Mix single component premixed reagent. When conducting experiments, the preparation of the PCR reaction solution is very convenient and simple, simply adding primers, probes, DNA or cDNA templates, and water. This product does not contain ROX correction dyes and is suitable for various fluorescent labeling probes, as well as for various quantitative PCR models.
The Fast Taq DNA polymerase in this kit uses proprietary hot start technology to ensure that there is no activity below 55 ℃, so a reaction system can be established at room temperature. Fast Taq DNA polymerase has excellent amplification speed, therefore it meets the requirements of fast PCR programs and can usually complete the amplification reaction within 30 minutes. At the same time, this polymerase has the ability to tolerate impurities and is also suitable for amplification reactions of crude nucleic acid samples.
Due to the extremely high amplification efficiency of PCR for nucleic acids, it is very easy to cause aerosol pollution in diagnostic PCR experiments, such as pipettes, countertops, PCR instruments, and air. When the experimental environment is contaminated, it can lead to false positive results for negative samples. In this reagent, all dTTPs in dNTP are replaced with dUTP, so that the amplified PCR product contains dU bases. In the next experiment, dU PCR products containing aerosol contamination will be degraded by the UNG glycosylase in this reagent (inactivated at 55 ℃ for 1 minute), thereby removing the aerosol contamination effect. The UNG enzyme in this reagent can eliminate up to 100000 copies of nucleic acid pollutants by incubating at 37 ℃ for 2 minutes. Therefore, this product is the best reagent for diagnostic quantitative PCR detection.
Kit features:
·The hot start enzyme adopts a proprietary modification method, with a 30 second hot start;
·Introducing UNG can efficiently eliminate aerosol pollution;
·Single component premixed solution, convenient and fast to use;
·Freeze dried products have extremely stable performance and are suitable for long-term storage;
·High sensitivity, capable of detecting low abundance genes;
·Suitable for all real-time quantitative PCR instruments.

Example of reaction:
Using the pUC57 plasmid as a template, perform 8 4-fold dilutions of the template, with a template copy number of 10 copies in the 8th dilution gradient/ μ l. Using this product for amplification testing, add template 2 to each reaction system μ l. As shown in the figure,
A: This product exhibits excellent linear relationship within a wide range of template quantities: amplification efficiency of 97.5%, R2=0.9995.
B: Test the ability of UNG to remove DNA contamination by adding 105 copies of dUTP containing PCR products to the reaction system for quantitative PCR. B1 is the freeze-dried probe fluorescence quantitative PCR kit, and B2 is the freeze-dried probe fluorescence quantitative PCR kit (UNG enzyme). From the figure, it can be seen that UNG can effectively remove 105 copies of aerosol pollution.
C: The standard curve of Figure A amplification.

Usage:
1. Dissolved freeze-dried products
Add 1ml of TaqMan PCR Buffer 7 to each bottle to dissolve the lyophilized product. The dissolved product is a TaqMan PCR Mix reagent (2 times the concentration) × TaqMan PCR Mix). The remaining reagents after dissolution can be stored at -20 ℃, and can be used directly after melting in the next experiment.
2. Configure reaction system
Prepare 20 according to the following components μ PCR reaction system

 

Prepare 20 according to the following components μ PCR reaction system  

Ingredients

Dosage

concentration

2×TaqMan PCR Mix(溶解后)

10μl

PCR Forward Primer (10μM)

0.4μl

0.2μM

PCR Reverse Primer (10μM)

0.4μl

0.2μM

TaqMan Probe (10μM)

0.4μl

0.2μM

cDNADNA

0.5~2μl

ddH2O

up to 20μl


3.  Perform Real Time PCR reaction using the following procedure:

Temperature

Time

Cycles

Explain

37℃

20min

1

Digestive pollutant

95℃

30s

1

Hot start reaction

95℃

5s

40

60

20s

 

 

 

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