Enzyme Labeling of Nucleic Acids

Enzyme Labeling of Nucleic Acids

Enzyme labeling of nucleic acids is a critical tool in molecular biology research. As a professional biotech company, BOC Sciences offers comprehensive and expert services in enzyme labeling of nucleic acids. Our highly skilled and experienced team is capable of working with a variety of enzymes and nucleic acids, ensuring the high quality and reliability of the labeling procedure.

What is Enzyme Labeling of Nucleic Acids?

Enzyme labeling of nucleic acids is a process of labeling nucleic acids (DNA or RNA) with enzymes in molecular biology. This process provides essential insights into gene expression, genome structure, and other key aspects of cellular function.

A common method of enzyme labeling is using labels such as fluorescent tags or radioactive isotopes that can be attached to an enzyme that binds to the nucleic acids. This allows for the tagged nucleic acid to be visualized, detected, or measured.

One of the widely used methods of enzymatic labeling is called nick translation, where the enzyme DNA polymerase makes a cut (nick) in the DNA strand, removes the piece of DNA where the nick was made, and inserts labeled nucleotides.

Another technique is random priming, which synthesizes short DNA fragments that complement the RNA or DNA sequences, which are subsequently incorporated into the newly synthesized DNA against the template.

Applications of Enzyme Labeling of Nucleic Acids

Enzyme labeling of nucleic acids is a technological advancement that has considerable implications in various scientific fields. Here are some applications primarily in the field of medicine and biological research:

Molecular Diagnostics: Enzyme labeling is extensively used in the detection and diagnosis of diseases. It can help healthcare professionals identify the presence of certain genes or genetic mutations that may lead to disease. In infectious diseases, enzyme-labeled probes help to detect specific pathogens by identifying their nucleic acids. For instance, it is used in the detection of HIV, hepatitis B, and other viruses.

Drug Discovery: Researchers use enzyme labeling to study the interaction between drugs and their molecular targets. This can help in the development of new, more effective drugs.

Genetic Research: Enzyme labeling is used in DNA microarrays to detect the presence and quantity of DNA sequences in a sample, which is a powerful tool for genomic research. It enables researchers to trace and identify specific DNA and RNA sequences.

Cancer Research: Enzyme labeling has revolutionized the field of oncology. It can be used to identify cancerous cells and to study the genetic changes that occur in cancer.

Food and Environmental Testing: It is also used to detect genetically modified organisms (GMO) in food products, or to track the presence of specific microorganisms in environmental samples.

Advantages of Enzyme Labeling of Nucleic Acids

High Sensitivity: Enzyme labeling of nucleic acids enhances the detection sensitivity. The enzyme attached to the probe can act on a substrate to generate a large amount of detectable products, significantly enhancing the signal, making it suitable for detecting small amounts of nucleic acids.

Specificity: Enzyme labeling allows the specific detection of particular sequences of nucleic acids, thus improving the accuracy and reliability of results.

Versatility: It allows a wide range of probes and detection systems to be used, making it adaptable for different research purposes.

Quantitative Analysis: It allows for quantitative analysis of DNA, RNA, and their interactions.

Real-Time Tracking: Enzyme labeling allows researchers to track the interactions of nucleic acids in real-time, which is crucial in understanding cellular processes and in drug discovery processes.

Less Hazardous: Radioactive labeling, another common method, uses radioactive isotopes which can pose health risks, whereas enzyme labeling is less hazardous.

Economical: As enzyme labeling does not rely on expensive and potentially dangerous isotopes, it is often a more cost-effective option.

Multiplexing: Enzyme labeling allows the simultaneous detection of multiple targets in the same sample, known as multiplexing, which is not possible with other methods.

Time-Efficient: It allows for faster detection and result generation which can speed up research processes.

Nucleic Acid Enzyme Labeling Services

Enzyme-Labeled DNA

Enzyme-labeled DNA involves attaching an enzyme to a DNA molecule. This labeling can be achieved through various methods, such as enzymatic conjugation or chemical modification. The enzyme attached to the DNA can then be used to catalyze specific reactions for detection or amplification purposes. For example, DNA molecules can be labeled with enzymes like horseradish peroxidase (HRP) or alkaline phosphatase (AP) to enable colorimetric or chemiluminescent detection methods in assays like ELISA (enzyme-linked immunosorbent assay) or PCR (polymerase chain reaction).

Enzyme-Labeled RNA

Similar to enzyme-labeled DNA, RNA molecules can also be conjugated with enzymes for various applications. Enzyme-labeled RNA is commonly used in techniques like RNA-based assays and in situ hybridization. For instance, RNA probes labeled with enzymes like digoxigenin or biotin can be used to detect specific RNA sequences in tissues or cells, allowing visualization of gene expression patterns.

Enzyme-Labeled Oligonucleotides

Oligonucleotides are short DNA or RNA molecules typically around 10-100 nucleotides in length. Enzyme-labeled oligonucleotides are designed for specific interactions with complementary nucleic acid sequences. Enzyme labels attached to oligonucleotides can be used for various applications such as detecting mutations, identifying specific sequences in samples, or facilitating enzymatic reactions like DNA or RNA cleavage.

Enzyme-Labeled Nucleotides

Enzyme-labeled nucleotides are commonly used in techniques like DNA sequencing, where they can be incorporated into newly synthesized DNA strands by DNA polymerases. The enzyme label allows for the visualization or detection of the incorporated nucleotides, aiding in the sequencing process.

Enzyme-labeled dNTPs: dNTPs are the building blocks of DNA, consisting of a nitrogenous base (adenine, guanine, cytosine, or thymine), a deoxyribose sugar, and three phosphate groups (triphosphate). Enzyme-labeled dNTPs are used in methods like PCR (Polymerase Chain Reaction) and DNA sequencing. These nucleotides can be labeled with various reporter molecules or enzymes (e.g., fluorescent dyes, biotin, or enzymes like polymerases) to allow for detection or visualization of DNA synthesis or extension events.

Enzyme-labeled ddNTPs: ddNTPs are modified nucleotides lacking the 3'-OH group on the sugar moiety, preventing further DNA chain elongation after their incorporation by DNA polymerase. Enzyme-labeled ddNTPs are essential components of Sanger sequencing, a method used for DNA sequencing. Each ddNTP is labeled with a different fluorescent dye or enzyme, allowing for base-specific termination of DNA synthesis during sequencing reactions.

Enzyme-Labeled Nucleosides

Enzyme-labeled nucleosides are nucleosides that have been conjugated with enzymes for specific applications, such as studying nucleoside metabolism or nucleoside-based signaling pathways.

Validation Services

We provide verification services for the labeling effect of nucleic acids, including labeling efficiency, labeling stability and other tests to ensure that the labeled nucleic acids can meet the needs of customers.

Professional Consulting Services

Provide professional consulting services and answer customers' questions about enzyme labeling.

Our Advantages

Extensive Experience: We possess vast experience in the field of biomedicine and particularly in Enzyme Labeling of Nucleic Acids.

Advanced Techniques: Our advanced technology and innovative techniques set us apart. We leverage state-of-the-art equipment and software in our processes.

Expert Team: Our team comprises experts with cutting-edge knowledge in biomedicine, biochemistry, and molecular biology.

High Success Rate: We have a broad portfolio detailing our high success rate and our proven methodologies in Enzyme Labeling of Nucleic Acids.

Customization: We offer customizable solutions to cater to each client's unique needs and requirements.

Quality Assurance: We adhere to stringent quality control measures to ensure accurate and reliable results.

Speedy Turnaround: We can promise a fast turnaround time for projects, aiming to aid our clients in their research or product development without unnecessary delays.

Cost-Efficiency: Our services are competitively priced, offering both quality and cost efficiency to our clientele.

Excellent Customer Service: We have a dedicated customer service team available to answer queries, deliver updates, and provide support to ensure our clients have the best experience with us.

Case Study

  • Case Study 1

The restriction endonucleases-assisted (REase-assisted) 3J probes were employed in combination with the electrochemical signal output. In this case, either a linear or hairpin probe/ reporter containing methylene blue as an electroactive label was immobilized on a gold electrode. In the presence of the target DNA, the 3J structure was formed, and the enzyme cleaved the reporter into two fragments. In the signal-OFF format, the methylene blue-labeled cleavage fragment of the reporter was released into solution along with the target. As a result, the peak current decreased in a target concentrationdependent manner. A detection limit of 14 pM was reported for the signal-OFF sensor with the linear reporter. The assay was able to differentiate between the target sequences differing in a single nucleotide. LOD of the hairpin capture probe was ~1 nM. In the signal-ON sensor design, the methylene blue label was attached to an internal thymidine of the reporter and was intercalated into its double-stranded region, which decreased the probability of the label to approach the electrode. The reporter cleavage released the electro-active label on the single-stranded fragment followed by the increase in the electrochemical signal. The disadvantages of the approach were poor LOD (~1 nM) and long assay time. Interestingly, out of six REases, most of which could work efficiently with 3J probes in a homogeneous format, only two enzymes -Bsp143I and FDSau3AI- were found to operate on solid support-bound 3J structures.

REase-assisted electrochemical DNA detection using 3J probes.Fig. 1 REase-assisted electrochemical DNA detection using 3J probes. (Gerasimova, Y. V., 2014)

  • Case Study 2

A polymerase ribozyme can be used to label the 3' end of RNA or DNA molecules by incorporating a variety of functionalized nucleotide analogs. Guided by a complementary template, the ribozyme adds a single nucleotide that may contain a fluorophore, biotin, azide or alkyne moiety, thus enabling the detection and/or capture of selectively labeled materials. The 24-3 polymerase ribozyme is a highly evolved form of the class I polymerase ribozyme that exhibits substantially less sequence preference compared to earlier forms of this enzyme. Employing either NTP or dNTP substrates, it is able to extend either an RNA or DNA primer on an RNA template to generate complementary RNA or DNA products. The polymerase contains 180 nt There is a short tag sequence at the 5' end of the polymerase that is complementary to a sequence at the 5' end of the template. Otherwise the template sequence is not constrained. The primer, which corresponds to the target nucleic acid, binds to the template through Watson-Crick pairing and is extended by the polymerase to achieve 3'-end labeling. In addition, four templates were constructed, each with a different templating nucleotide at the first position of primer extension, followed by several non-complementary nucleotides (Figure 1B). Self-complementary and oligo(G) sequences were avoided within the non-complementary region to prevent formation of secondary structure. Together this set of templates enabled the testing of NTP analogs containing each of the four nucleobases.

3-End labeling of nucleic acids by the 24-3 polymerase ribozyme.Fig. 2 3'-End labeling of nucleic acids by the 24-3 polymerase ribozyme. (Samanta, B., 2018)

FAQ

1. How long does the enzyme labeling of nucleic acids service typically take?

The time frame for the enzyme labeling of nucleic acids service can vary depending on several factors, including the complexity of the project and the quantity of the sample.

2. What types of nucleic acids can be labeled?

Our service is capable of labeling various types of nucleic acids, from DNA and RNA to more complex molecules.

3. How accurate is the enzyme labeling of nucleic acids service?

We strive for the highest degree of accuracy in our enzyme labeling service. However, the accuracy can be influenced by various factors including the quality of the initial sample.

4. What is the cost of this service?

The cost of the enzyme labeling of nucleic acids service depends on various factors, including the complexity and scale of the project.

5. Can you provide this service for large scale projects?

Yes, we have the capability to handle both small and large scale projects.

6. How can I prepare my samples for enzyme labeling?

We provide detailed instructions on sample preparation to ensure optimal results.

7. How are the results delivered?

The results are typically delivered in a report that includes detailed analysis and data interpretation.

References

  1. Gerasimova, Y. V., et al. Enzyme-assisted target recycling (EATR) for nucleic acid detection. Chemical Society Reviews. 2014, 43(17): 6405-6438.
  2. Samanta, B., et al. 3'-End labeling of nucleic acids by a polymerase ribozyme. Nucleic Acids Research.2018, 46(17): e103-e103.
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