<i>Evo M-MLV</i> 一步法 RT-qPCR 预混液 （适用于猪蓝耳病毒检测， 含 UNG，不含引物探针)

Evo M-MLV One-step RT-qPCR premix (for porcine blue ear virus, contains UNG, no primer probe)

Product Code: AG11755

- +

¥3,600

Product Description

This is a 2X Premix Probe Kit for the detection of porcine blue ear virus (PRRSV). Reverse transcription and qPCR are done in the same reaction system, and the reaction solution is very simple and quick to prepare, only primers, probes, templates, and RNase free water need to be added to carry out the qPCR reaction, which can effectively reduce the risk of contamination.
The Evo M-MLV reverse transcriptase and Accurate Taq HS DNA polymerase with excellent reaction performance, together with carefully optimized Buffer, can synthesize cDNA and carry out highly efficient and stable qPCR amplification in a short period of time, which is very suitable for the detection of swine blue ear and other RNA viruses.
The dUTP/UNG Anti-Contamination System is introduced in this product, which replaces dTTP with dUTP in the PCR reaction, and utilizes UNG enzyme to selectively hydrolyze dU-containing DNA strands without affecting non-dU-containing DNA strands, thus removing dU-containing contaminating templates introduced during the preparation of the PCR reaction system, and thus preventing the generation of false-positive results of the PCR effectively; At the same time, this product is mixed with a monoclonal antibody that can inhibit the activity of DNA Polymerase at room temperature, 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 one-step RT-qPCR premixed kit, the preparation of reaction solution is very simple and fast, only need to add primers, probes, templates and RNase free water to carry out the qPCR reaction, which effectively reduces the possibility of contamination due to multiple operations.
2. This product adopts reverse transcriptase and hot-start polymerase with excellent reaction performance, together with the carefully optimized reaction system, featuring strong specificity and high amplification efficiency, which is very suitable for the detection of RNA viruses such as porcine blue ear.
3. dUTP/UNG is a contamination prevention system that removes contamination of dU-containing PCR products to prevent false-positive PCR results and improve the accuracy of the results.

Product Composition
individual parts making up a compound norm
2X One Step RT-qPCR Probe Premix for PRRSV (UNG Plus, Primer Free)*1 1 ml X 5 pcs
RNase free water 1 ml X 5 pcs

*1: Contains Evo M-MLV RTase Enzyme, Accurate Taq HS DNA Polymerase, UNG Enzyme, RNase Inhibitor, dNTP Mixture and reaction buffer.

Preservation and transportation
Storage temperature: -20℃ storage
Transportation temperature:Dry ice transportation or -20°C ice pack transportation.
Example 1

Using the porcine blue-ear virus PRRSV JXA1 standard [ GBW(E)090929 ] as template, this product was used for the detection of O gene (FAM) and NSD9 gene (HEX) by fluorescent probe qPCR method, and the amount of template in the reaction system was 250 copies, 50 copies, 10 copies, 2 copies, 0.4 copies, 0.5 copies, 0.5 copies, 0.5 copies, 0.5 copies and 0.5 copies. The amount of template added in the reaction system was 250 copies, 50 copies, 10 copies, 2 copies, 0.4 copies and 0 copies.
Quantification instrument used: ABI QuantStudioTM 5 Real-Time PCR Systems, and the results of the experiments were as follows:

The results are shown above:

1, different concentrations of templates can get good amplification within 45 cycles;
2. Negative controls were not detected within 45 cycles;
3. The amplification efficiency was 97%, R2=0.999.


The results are shown above:

1, different concentrations of templates can get good amplification within 45 cycles;
2. Negative controls were not detected within 45 cycles;
3. The amplification efficiency was 95%, R2=0.993.

 

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