Hifair™ Superior Universal Multiplex One Step RT-qPCR Probe Kit (UDG Plus) _ 16901ES (2G)

Hifair™ Superior Universal Multiplex One Step RT-qPCR Probe Kit (UDG Plus) is a multiplex quantitative PCR kit based on RNA as template. In the process of the experiment, reverse transcription and quantitative PCR were carried out in the same tube, which simplified the experimental operation and reduced the risk of contamination.

In this kit, the first strand cDNA was efficiently synthesized by heat-resistant Hifair™ Reverse Transcriptase and quantitatively amplified by UNICON™ HotStart Taq DNA Polymerase. The kit mainly contains optimized MP buffer, enzymes mix. The buffer solution already contains Mg²⁺ and dNTP Mix. In addition, the factors that can effectively inhibit the non-specific PCR amplification and improve the amplification efficiency of multiple qPCR reactions are added, which can ensure the amplification efficiency and carry out up to multiple amplification reaction. The dUTP/UDG system was added to effectively prevent the risk of aerosol contamination.

Feature

Features

1. Good versatility: It is suitable for qPCR amplification and detection of nucleic acids of pathogenic microorganisms, human genomes, plants and other species.

2. High sensitivity: The carefully optimized buffer system improves the detection sensitivity of low-concentration templates up to 250 copies/mL.

3. dUP/UDG system: dUTP/ Heat-labile UDG anti-pollution system is introduced to degrade aerosol pollution of products efficiently, reduce false positive, and ensure authentic results.

4. Supporting fast program: compatible with 40min fast program.

5. Super stability: stable at 37℃ for 14 days,repeated freezing and thawing 10 times.

Specifications

Components

Note: 1) 5×Hifair™ MP Buffer is the abbreviation of Hifair™ Superior Multiplex One Step RT-qPCR Probe Buffer, which includes dNTPs, dUTP, Mg²⁺, stabilizers and more.

2) Hifair™ Enzyme Mix mainly contains Hifair™ reverse transcriptase, UNICON™ HotStart Taq DNA polymeras, RNaseinhibitor, and UDGase and so on.

Storage

The product should be stored at -25℃ ~ -15℃ for 1 year. It should avoid repeated freezing and thawing.

Instructions

1. Reaction Composition:

Note：Be sure to mix well before use, avoid excessive bubbles caused by violent vibration.

a) Primer concentration: Primer mix including multiplex primer, depending on the situation optimal primer concentration may be between 0.1 and 1.0 μM.

b) Probe concentration: Probe mix including multiplex probe labeling difference fluorescent group, depending on the situation optimal probe concentration may be between 0.05 and 0.5 μM.

c) Template dilution: qPCR is highly sensitive and it is recommended to dilute the template. The control Ct value is suitable between 20 and 35.

d) System preparation: un head with filter element. Avoid cross contamination and aerosol contamination.

2. Optimized Cycling Protocol

(1) Standard amplification procedure

Note：

a) Reverse transcription: The temperature can select 50°C or 55°C for 10-15 minutes.

b) Amplification reaction: The temperature is adjusted according to the Tm value of the designed primers.

c) Fluorescence signal acquisition: Please set the experimental procedure according to the requirements of the instrument manual.

(2) Rapid amplification program

Note：If the actual qPCR detector supports rapid amplification, please perform a pre-test to confirm.

3. Application equipment

ABI 5700, 7000, 7300, 7700, 7900HT Fast, StepOne™, StepOne Plus™，ABI 7500, 7500 Fast, ViiA™7, QuantStudio™ 3 and 5, QuantStudio™ 6,7,12k Flex；

Stratagene MX3000P™, MX3005P™, MX4000P™；

Bio-Rad CFX96™, CFX384™, iCycler iQ™, iQ™5, MyiQ™, MiniOpticon™, Opticon®, Opticon® 2, Chromo4™；

Eppendorf Mastercycler® ep realplex, realplex 2 s;

Qiagen Corbett Rotor-Gene® Q, Rotor-Gene® 3000, Rotor-Gene® 6000；

Roche Applied Science LightCycler® 480, LightCycler® 2.0; Lightcycler® 96；

Thermo Scientific PikoReal Cycler; Cepheid SmartCycler®; Illumina Eco qPCR; SLAN 96S, 96P.

Notes

Please wear the necessary PPE, such lab coat and gloves, to ensure your health and safety！

Please use RNasefree consumables for the experiment.

This product is for research use only！