2X 预混型探针法 qPCR 试剂盒 II（含 UNG）

2X Premixed Probe-based qPCR Kit II (with UNG)

Product Code: CM0213

- +

¥400

Product Description

This product is a 2X Premix premix that is easy to use and requires only the addition of primers, probes, templates, and water to perform the reaction.
This product contains the superior reaction performance of Accurate Taq HS DNA polymerase, together with carefully optimized reaction buffer, improves the PCR amplification performance and sensitivity of low concentration template detection, enabling highly sensitive Real Time qPCR reactions for accurate quantitative detection of target genes, and supporting pre-mixing of primer probes.
The dUTP/UNG anti-contamination system is introduced in this product. dUTP replaces dTTP in the PCR reaction, and the UNG enzyme selectively hydrolyzes dU-containing DNA strands without affecting non-dU-containing DNA strands, thus removing dU-containing contaminating templates introduced in the preparation of the PCR reaction system, and thus preventing the generation of false-positive results in the PCR reaction effectively; At the same time, monoclonal antibody that can inhibit the activity of DNA Polymerase at room temperature is also added to this product, which can effectively inhibit non-specific amplification, improve the sensitivity of the reaction and enhance the accuracy of the results.

 

Product Advantages

1. This product is a 2X premix, the reaction solution is very simple to prepare, only need to add primers, templates, probes and water for qPCR reaction.
2. This product utilizes a superior performance Accurate Taq HS DNA polymerase and optimized reaction system with high specificity and amplification efficiency.
3. This product supports premixing of primer probes.
4. The dUTP/UNG contamination prevention system has been introduced into the product to eliminate contamination of dU-containing PCR products, thereby preventing false positive PCR results and improving the accuracy of the results.

Product Composition
individual parts making up a compound norm
2X Accurate Taq HS Probe Premix II (UNG Plus) 1 ml

 

Preservation and transportation
Storage temperature: -20℃ storage
Transportation temperature: dry ice transportation or -20℃ ice bag transportation
Example 1

Pre-mixed primer probes Pre-mixed primer probes Stability accelerated at 37 °C for 5 days
Human cDNA (100 ng ~ 1 pg) was used as template, RNase free water was used as negative control instead of template amount, primer probes were pre-mixed, and then accelerated and untreated samples were subjected to triple assay at the same time after being placed at 37℃ for 5 days to test the performance of the TFR gene (GC content 38%, FAM channel), Actin gene (GC content 59%, HEX channel), and RPP30 gene (GC content 62%, Cy5 channel). TFR gene (GC level 38%, FAM channel), Actin gene (GC level 59%, HEX channel), and RPP30 gene (GC level 62%, Cy5 channel) were tested simultaneously in a triplex assay for the TFR gene (GC level 38%, FAM channel), Actin gene (GC level 59%, HEX channel), and RPP30 gene (GC level 62%, Cy5 channel).
The quantitative instrument used for the assay was the ABI QuantStudio.TM 5 Real-Time PCR Systems, the results of the triple test are shown below:


The results are shown above:
1、Accurate quantification can be carried out in a wide range of templates, and the amplification curves of qPCR reactions with 100 ng ~ 1 pg cDNA show good linearity.;
2. Negative controls were not detected within 45 cycles.;
3. After pre-mixing the primer probes and placing them at 37℃ for 5 days, the performance of amplifying the gene was comparable to that of untreated samples.


The results are shown above:
1、Accurate quantification can be carried out in a wide range of templates, and the amplification curves of qPCR reactions with 100 ng ~ 1 pg cDNA show good linearity;
2. Negative controls were not detected within 45 cycles.;
3. After pre-mixing the primer probes and placing them at 37℃ for 5 days, the performance of amplifying the gene was comparable to that of untreated samples.


The results are shown above:
1、Accurate quantification can be carried out in a wide range of templates, and the amplification curves of qPCR reactions with 100 ng ~ 1 pg cDNA show good linearity;
2. Negative controls were not detected within 45 cycles.;
3. After pre-mixing the primer probes and placing them at 37℃ for 5 days, the performance of amplifying the gene was comparable to that of untreated samples.

Example 2

Support for fast programs

The African swine fever B646L standard material [ GBW ( E ) 091034 ] was used as the template, and the fluorescent probe qPCR method was used to detect the B646L gene, and the amount of template added in the 25 μl reaction system was 35 copies and 0 copies (negative control). The standard and rapid procedures were performed after the preparation of the reaction system, and the quantification instrument used was ABI QuantStudio.TM 5 Real-Time PCR Systems. the program is as follows:

The results of the experiment are as follows:

The results are shown above:
1. For low-copy virus templates, the difference between the mean ct values of different replicate wells in the standard mode and the fast mode is △ct < 0.5, and the out-put rates are comparable;
2. Negative controls were not detected within 45 cycles.

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