Next-generation Polymerase Chain Reaction (PCR)

With the development of biomolecular fluorescence technology, the second-generation PCR (qPCR) based on fluorescent probes or dyes has gradually become the mainstream molecular diagnostic technology for detecting nucleic acid fragments. However, PCR and qPCR cannot achieve absolute quantification of samples.

Fig.1 Comparison of PCR, qPCR, and dPCR technologies. (Quan, et al., 2018)

With the continuous maturity of microelectromechanical systems and the development of microfluidic technology, a new-generation PCR, that is, digital PCR, appears. dPCR is a kind of absolute quantitative PCR that first dilutes the PCR reactants, then amplifies in different reaction chambers, and finally obtains the initial copy number or concentration of the molecule by Poisson's statistics.

Fig.2 The fundamentals of dPCR. (Quan, et al., 2018)

dPCR-based Glycogene Expression Profiling at CD BioGlyco

CD BioGlyco has mastered cutting-edge dPCR technology and has various dPCR platforms, such as active partitioning, passive partitioning, self-digitization, and droplet-based platforms. We aim to achieve the optimum performance of dPCR by providing a high number of partitions with limited volume variations and a large total reaction volume.

Fig.3 Different dPCR platforms to achieve massively parallel sample partitioning. (CD BioGlyco)

With dPCR-based glycogene expression profiling provided by CD BioGlyco, clients realize any idea:

• Detection of rare mutated glycogenes: Compared to wild-type DNA, the proportion of cancer-associated mutated sequences is very low, and is often undetectable by ordinary qPCR. The dPCR technology provided by CD BioGlyco, with its ultra-high sensitivity, easily quantifies mutation frequencies up to 0.0001%. We accordingly provide various Cancer Glycogene Discovery Services.
• Detection of pathogenic microorganisms: Analysis of accurate viral load is essential to elucidating the disease course, subsequent treatment options, and efficacy evaluation. In the study of pathogens, it is very important to quantify the changes of viral glycogenes accurately and reliably, as even small fluctuations in viral load are significant. CD BioGlyco has a deep knowledge of dPCR and helps clients easily achieve their goals.
• Analysis of glycogene expression differences: qPCR generally only detects a greater degree (more than twice) of difference, while the detection accuracy of dPCR exceeds ±10%, which easily identifies smaller differences in glycogene expression.
• Seamless integration with Next-Generation Sequencing (NGS): Compared with NGS, dPCR enriches the target glycogens in the sequenced samples, as well as verifies and accurately quantifies the sequencing results. The combination of these two technologies helps researchers obtain more accurate and reliable glycogene expression profile data.

Applications

• Detection of rare glycogene mutations
• Detection of pathogenic microorganisms
• Analysis of glycogene expression differences
• Identification of potential drug targets and development of targeted therapeutic drugs
• Development of personalized diagnosis and therapy for tumors
• Development of precise molecular diagnostic methods

Highlights

• Extremely high sensitivity: Single nucleic acid molecules are detected with detection limits as low as 0.001%
• No standard curve required
• Suitable for the detection of samples with complex substrates
• Effective differentiation of samples with small concentration differences

CD BioGlyco has developed an advanced Glycogenomics Platform and has been widely praised by customers around the world. We adhere to the service-paramount concept, aiming to serve every client in the field of biotechnology with advanced technology and an excellent platform. Please feel free to contact us for more detailed information.