Benefits

 

• Precise and accurate automatic process saves tremendous time for sample preparation and improves test efficiency
• High compatibility with most consumables eliminates the need for replacing lab ware in the test workflow

 

Overview

 

Amid the widespread outbreak of the COVID-19 virus, the development of test methods that can efficiently and precisely identify positive carriers is cited as one of the most important approaches to curbing the pandemic, other than the development of vaccines and medications. Among the known methods for testing virus, the qPCR test is used most widely, while a new method developed by Israeli researchers at Technion University, which is based on qPCR and pool testing and can test a batch of pooled samples at once for high volume screening, is gaining popularity among laboratories for COVID-19 testing. However, these methods involve skillful operations and tedious processes that could pose a serious challenge to lab staff.

 

The EzMate Automated Pipetting System was designed to simplify the pipetting process and eliminate errors resulting from manual operations. When used for sample preparations in a conventional qPCR practice, the EzMate, as compatible with most consumables in the lab supplies market, can seamlessly fit into the process and automatically take samples from a nucleic acid extraction system and implement pre-PCR processing before loading the plate onto the qPCR machine. No replacement of consumables is needed in the process, thus decreasing the potential of cross contamination and speeding up sample preparations.

 

In addition, when a pooling method is applied in qPCR testing, the EzMate can also be used in sample preparations to provide automated pipetting in place of traditional manual practices, so as to streamline sample preparation workflow, avoid errors and improve test throughput.

 

In the following example, we’ll demonstrate how the EzMate can be applied in the practices of qPCR testing with/without pooling samples for COVID-19 screening, and you’ll see how the machine can help improve lab throughput.

 

Workflow

 

The overall workflow is described in figure 1 and separated into two processes if pooling is necessary. The compatible consumables are described as figure 2.

 

 

Figure 1. Workflow of qPCR detection by using EzMate automated pipetting system.

 

After sampling, EzMate automated pipetting system helps pooling, nucleic acid extraction, and qPCR sample preparation for qPCR detection. It saves manpower,  time and avoids human errors to improve accuracy and precision.

 

Figure 2. Compatible with a variety of consumables.

A variety of consumables being compatible with EzMate automated pipetting system makes users convenient to choose consumables.

 

A-1. Sample preparation in plate from nucleic acid extraction system (Figure 3):

 

  1. Sampling

  2. Nucleic acid extraction

  3. qPCR sample preparation

       a. Transfer mastermix

       b. Transfer sample and control

  4. qPCR detection

 

(A) Mastermix is prepared and separated to 4 tubes (big blue rounds) and transferred to a 96 well PCR plate (red rounds). Nucleic acids of samples are extracted in the deep well plate (small blue rounds) and transferred to the 96 well PCR plate (red rounds). After preparation, the plate is ready for qPCR detection. Green rounds indicate discarded tips used in sample preparation. N and P indicate negative and positive control, respectively. Only mastermix preparation and separation is made by hand. Others are automated.

(B) Adaptors and pipetting module package needed for preparation in plate.

 

(A)

(B)

Figure 3. Direct sample preparation in plate for qPCR detection from nucleic acid extraction system.

 

A-2. Sample preparation in tube from nucleic acid extraction system(Figure 4):

 

(A) Mastermix is prepared and separated to 4 tubes (blue rounds in left panel) and transferred to a 96 well PCR plate (red rounds). Nucleic acids of samples are extracted in sample tubes (blue rounds in middle panel) and transferred to the 96 well PCR plate (red round). After preparation, the plate is ready for qPCR detection. Green rounds indicate discarded tips used in sample preparation. N and P indicate negative and positive control, respectively (purple rounds). Only mastermix preparation and separation is made by hand. Others are automated.

 

(B)Adaptors and pipetting module package needed for preparation in tube.

 

(A)

(B)

B. Sample Pooling before QPCR detection (Figure 5 and 6):

 

  1. Sampling

  2. Transfer sample

  3. Pooling (P-BEST pooling method is described by Hertz et al., 2020*)

  4. Nucleic acid extraction

  5. qPCR sample preparation

       a. Transfer mastermix

       b. Transfer sample and control

  6. qPCR detection

 

 (A)

(B)

Figure 5. Sample transferring.

 

(A) Samples are first placed in tubes (blue rounds) and then transferred to a 96 well PCR plate (red rounds) automatically. Green rounds indicate discarded tips used in sample preparation.

(B) Adaptors and pipetting module package needed for sample transferring.

 

(A)

(B)

(A) For P-BSET pooling method, there are 384 samples prepared for pooling (blue rounds). The 384 samples are transferred from 96 well PCR plates to 48 tubes automatically with a program code writtenwith Python to complete pooling (Hertz et al., 2020). After pooling, the 48 tubes are prepared for nucleic acid extraction as described in figure 4.

(B) Adaptors and pipetting module package needed for P-BEST pooling.

 

Note:*

Shental, N., Levy, S., Wuvshet, V., Skorniakov, S., Shalem, B., Ottolenghi, A., Greenshpan, Y., Steinberg, R., Edri, A., Gillis, R., Goldhirsh, M., Moscovici, K., Sachren, S., Friedman, L.M., Nesher, L., Shemer-Avni, Y., Porgador, A., and Hertz, T. (2020). Efficient high-throughput SARS-CoV-2 testing to detect asymptomatic carriers. Sci. Adv. 6: eabc5961.

 

DOWNLOAD APPLICATION NOTE

 

PCR/qPCR Set Up Related Applications

 

• Evaluate the Performance of Real-time PCR Instruments by Using EzMate 401 Automated Pipetting System
• Accelerate qPCR Setup for COVID-19 Diagnosis on EzMate 601s Automated Pipetting System

---

 

Learn More About EzMate Automated Pipetting System

 

We offer customized solutions to suit your experiment need, contact us for further discussion.