Extraction and PCR Services

Empowering Your Genetic Discoveries with the Highest Accuracy

Polymerase Chain Reaction (PCR) is a fundamental technique in molecular biology, widely used for amplifying specific DNA sequences. PCR typically requires DNA and/or RNA extraction which is performed prior to sequencing or PCR amplification. PCR is typically performed after sequencing or during the experiment to troubleshoot results.

Central to the success of PCR are oligonucleotides, which serve as primers and probes, guiding the amplification process with precision and specificity. Despite this connection, many oligonucleotide manufacturers are limited in their PCR capabilities due to the possibility of cross contamination when synthesizing oligos in close proximity of PCR amplification. Eurofins Genomics is unique in this way. We have two separate facilities located adjacent to each other, which completely separates oligonucleotide manufacturing from PCR and sequencing. This allows us to safely synthesize primers and probes to use in PCR assays and run the PCR services as well.

The PCR process typically begins with extraction. Through meticulous methodologies, we extract high-quality DNA or RNA from various sample types, including blood, cells, viruses, and waste. Our skilled scientists leave no room for contaminants, safeguarding the integrity of your DNA and providing you with trustworthy data to fuel your discoveries.

Sample Types Accepted

Blood		Saliva		Environmental		Mammalian		Virus

Why Eurofins?

• Full Service Provider: We are one of the only labs in the country that can safely synthesize custom designed assays and run PCR experiments from the same location, by having side-by-side facilities.
  
• Scientific Excellence: Our team comprises distinguished scientists with a passion for precision in genetic research.
• Customized Solutions: Tailored to your unique research requirements, ensuring optimal outcomes for your projects.
• Data Integrity and Security: Trust in our robust quality control measures, safeguarding the confidentiality of your data.
• Timely Delivery: We understand the value of time in research, and our streamlined processes ensure timely delivery of results.

Industries We Serve

		Academia and Research Elevate your academic pursuits with reliable and validated genetic data, shaping the future of scientific exploration.
		Pharmaceuticals and Biotechnology Drive drug discovery and development with precise genetic information, revolutionizing patient care and biotechnological solutions.
		Environmental Sciences Unravel the secrets of biodiversity and ecological dynamics, influencing conservation efforts and environmental understanding.
		Agriculture and Crop Improvement Harness the power of genetics to improve crop breeding programs, ensuring sustainable and high-yield agricultural practices.

Contact Us

Our staff are happy to answer any questions that you may have. Email us at GenomicsSupport@eurofins.com to discuss the needs of your project.

The Role of Oligonucleotides in PCR Research

Oligonucleotides are short sequences of nucleotides, typically ranging from 15 to 30 bases in length. In PCR, they function primarily as primers, which are essential for initiating DNA synthesis. The two primers, known as forward and reverse primers, are designed to be complementary to the sequences flanking the target region of DNA. During PCR, these primers anneal to their complementary sequences on the template DNA, providing a starting point for DNA polymerase to extend the DNA strand.

Primer Design: Precision and Specificity

The design of oligonucleotide primers is a critical aspect of PCR. Successful amplification depends on primers that are specific to the target sequence and do not bind to non-target regions. Several factors are considered in primer design, including:

• Length: Primers are typically between 18 and 25 nucleotides long, providing a balance between specificity and efficient annealing.
• Melting Temperature (Tm): The Tm of the primers should be similar, usually within 2-5°C of each other, to ensure simultaneous annealing during PCR cycles.
• GC Content: A balanced GC content (typically 40-60%) helps maintain stable primer binding.
• Secondary Structure: Primers should be designed to avoid the formation of secondary structures such as hairpins or dimers, which can interfere with amplification.

Oligonucleotides as Probes in Real-Time PCR

In addition to serving as primers, oligonucleotides are also used as probes in real-time PCR (qPCR). Probes are short, sequence-specific oligonucleotides labeled with a fluorescent reporter dye. They hybridize to a specific sequence within the PCR product, allowing for real-time detection and quantification of DNA as it is amplified.

Common types of probes used in qPCR include:

• TaqMan Probes: These probes contain a fluorescent reporter dye at one end and a quencher at the other. During PCR, the probe is cleaved by the 5' to 3' exonuclease activity of Taq polymerase, separating the reporter from the quencher and resulting in fluorescence that correlates with the amount of amplified product.
• Molecular Beacons: These probes form a hairpin structure with a reporter and quencher in close proximity. Upon hybridization to the target sequence, the hairpin opens, separating the quencher from the reporter and producing a fluorescent signal.
• Scorpion Probes: These probes combine the properties of primers and probes in a single molecule. They hybridize to the target sequence and generate fluorescence during PCR.

Applications of Oligonucleotides in PCR Research

Oligonucleotides are indispensable in a wide range of PCR-based applications, including:

• Gene Expression Analysis: Real-time PCR allows for the quantification of gene expression levels by amplifying cDNA generated from mRNA. Oligonucleotide probes and primers ensure specific detection of target transcripts.
• Genotyping and Mutation Detection: PCR is used to identify single nucleotide polymorphisms (SNPs) and other genetic variations. Allele-specific primers and probes enable the detection of specific mutations.
• Pathogen Detection: Oligonucleotide primers and probes are used in diagnostic PCR assays to detect the presence of viral or bacterial DNA in clinical samples, providing a rapid and sensitive method for identifying pathogens.
• DNA Sequencing and Cloning: PCR is a precursor to DNA sequencing and cloning, where oligonucleotide primers amplify specific DNA regions, which are then sequenced or inserted into vectors for further study.

Advancements in Oligonucleotide Technology

Recent advancements in oligonucleotide synthesis and modification have further enhanced the utility of oligonucleotides in PCR. Locked Nucleic Acids (LNA), Peptide Nucleic Acids (PNA), and other modified nucleotides improve the stability, specificity, and binding affinity of oligonucleotides, allowing for more accurate and robust PCR results.

Oligonucleotides are at the heart of PCR research, enabling the precise amplification and detection of DNA sequences. Their design and application are critical to the success of PCR in various fields, from basic research to clinical diagnostics. As PCR technology continues to evolve, the role of oligonucleotides remains central, driving innovations in molecular biology and biotechnology.