COVID-19 nucleic acid detection raw material solution: fast, freeze-dried

At present, the SARS-CoV-2 coronavirus that causes COVID-19 pandemic has mutated over time, resulting in spreading faster and more stealthily. The prevention and control of pandemics should be deeply concerned in this critical situation. Yeasen Biotechnology (Shanghai) Co., Ltd. continuously provides customers with high-quality and stable nucleic acid detection raw materials to speed up and simplify COVID-19 nucleic acid testing.

Solution 1: Extraction-free rapid RT-qPCR, complete detection in 30 minutes

Product collocation: 

Nucleic Acid Release Reagent (Cat#13656ES) (Inquire)

Hieff Unicon™ Fast Multiplex One Step RT-qPCR Probe Kit(UDG plus) (Cat#13660ES)(Inquire)

1/ Nucleic Acid Release Reagent (13656ES)(Inquire)

One-step extraction-free nucleic acid release agent, suitable for rapid lysis of RNA templates from serum, plasma, bronchoalveolar lavage fluid and swab samples, without heating or nucleic acid extraction/purification process. Add this reagent to the sample when using and can extract RNA at room temperature in 10 min.

Features:

1 single solution, easy to operate, efficient and fast;

2 Compatible with various follow-up experiments, including qPCR, RT-qPCR, etc.;

3 The reagent is safe, toxic-free, environmentally friendly and healthy;

4 The product can be shipped and stored at room temperature;

5 It can be used as a sample preservative to delay RNA degradation.

2/ Hieff Unicon™ Fast Multiplex One Step RT-qPCR Probe Kit(UDG plus) (Cat#13660ES)(Inquire)

Designed for virus samples, virus detection can be quickly completed with only one tube of reaction solution. This one-step method is suitable for fast cycling reactions, and experimental results can be obtained within 30 min.

Features:

1 30 min rapid amplification, suitable for rapid amplification procedures of various instruments such as Q5 and SLAN;

2 Good reverse transcription specificity and extension performance;

3 High amplification efficiency, high amplification sensitivity, and high amplification specificity;

4 dUTP/UDG anti-pollution system.

Case show:

1) 10 μL of pseudovirus RNA was obtained by using nucleic acid release agents from Yeasen and S* respectively. After the samples were stored for 4 hours, they were amplified and verified with RT-qPCR reagents. The results show that the nucleic acid obtained by Yeasen nucleic acid release agent has better sensitivity and amplification linearity when matched with one-step RT-qPCR reagent.

Figure 1. Extraction-free pseudovirus amplification curve

Amplification Instrument and Procedure: Q5 Fast

Yellow: water; Blue: s*; Red: Yeasen

2) The virus sample storage time test of Yeasen nucleic acid release agent was verified by amplification with the RT-qPCR reagent. It was found that the amplification effect was still good after the samples were placed for 34 days.

Figure 2. Virus RT-qPCR amplification curve under different treatment time of release agent

Amplification Instrument and Procedure: Q5 Fast

Solution 2: Freeze-dried RT-qPCR, effectively reducing costs

Traditional molecular diagnostic reagents have problems in shipping and storage. In order to ensure the biological activity of the active ingredients (DNase, reverse transcriptase, etc.) in the reagent, it is generally necessary to store and transport in a cold chain environment around -20°C. Cold chain transportation requires a lot of ice packs or dry ice, which is costly, and changes in ambient temperature can easily cause repeated freezing and thawing of detection reagents, which will directly affect the performance and shelf life of the reagents. Lyophilization of liquid reagents into solid reagents will solve the above problems.

Product collocation: 

Hieff Unicon™ V Lyo-nCoV Multiplex One Step RT-qPCR Kit(with MgCl2) (CAT#13775)

qPCR Lyoprotectant (CAT#13743)

1/ Hieff Unicon™ V Lyo-nCoV Multiplex One Step RT-qPCR Kit(with MgCl2) (CAT#13775)

A glycerol-free, ready-to-use molecular diagnostic reagent, containing thermostable reverse transcriptase, hot-start Taq DNA polymerase, reaction buffer, dNTPs, magnesium chloride, and lyoprotectant. The reagent is also added factors effectively inhibiting non-specific PCR amplification and factors improving the amplification efficiency of multiplex qPCR reactions. This product is an ideal choice for the development of multiplex RT-qPCR that has room temperature stability and can be shipped and stored at room temperature.

Advantages:

👍1 After lyophilization, the enzyme activity remains intact, and the performance is the same as that of liquid reagents;

👍2 After lyophilization, the product remains stable at room temperature, and does not require cold chain transportation and low-temperature storage;

👍3 Reduce cost and operational complexity: Provide reference freeze-drying process to help lyophilization process; no additional freeze-drying protective agent is required, and it can be used directly for lyophilization.

Partial data display

Figure 3. Liquid performance of 13775: Multiplex RT-qPCR amplification using high-sensitivity RT-qPCR reagents 13110 (red) and 13775 (blue) for pseudovirus templates. The left graph is FAM channel, the right is VIC channel.

4. Thermal stability of 13775 liquid reagent at 37°C for 5 days: The 13775 liquid reagent was placed at 37°C (green) and -20°C (blue) for 5 days for multiplex RT-qPCR amplification test. The left graph is FAM channel, the right is VIC channel.

Figure 5. The performance of 13775 product after lyophilization: the pseudovirus template was amplified by multiple RT-qPCR using liquid reagents (red) and 13775 lyophilized powder (purple) respectively. The left graph is FAM channel, the right is VIC channel. The results showed that the activity of 13775 reagent was intact after lyophilization, and it still had high-efficiency multiple reaction ability.

Figure 6. Thermal stability of 13775 lyophilized powder—37°C for 21 Days: The 13775 lyophilized powder was placed at 37°C (green) and -20°C (blue) for 21 days for multiple RT-qPCR amplification tests. The left graph is FAM channel, the right is VIC channel. The results showed that the lyophilized powder still had a good amplification effect after being placed at 37°C for 21 days.

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