Understanding Solid Sequencing Technology and Its Key Enzyme

The enzyme used in Solid sequencing technology is :
A. Sequenase
B. DNA polymerase
C. DNA Ligase
D. Taq Polymerase

Solid sequencing technology is a high-throughput method for sequencing DNA, widely used in genomics and molecular biology research. This technique relies on a specific enzyme to facilitate the sequencing process. For students preparing for CSIR NET Life Science, IIT JAM, GATE Biotechnology, and DBT JRF, understanding the key components and mechanisms of solid sequencing is essential for mastering molecular biology.

Correct Answer: (C) DNA Ligase

The correct answer is (C) DNA Ligase because solid sequencing technology uses DNA ligase to join nucleotide probes during the sequencing process. This enzyme plays a crucial role in the accuracy and efficiency of sequencing.

Keyphrase: Solid Sequencing Technology

Solid sequencing technology is a next-generation sequencing (NGS) method that uses DNA ligase to detect and identify nucleotide sequences accurately.

What is Solid Sequencing Technology?

Solid sequencing (Sequencing by Oligonucleotide Ligation and Detection) is an advanced next-generation sequencing (NGS) method. It involves the hybridization of short oligonucleotide probes to a single-stranded DNA template, followed by ligation using DNA ligase. The sequence is determined based on the specificity of the probes and the order in which they are ligated.

Mechanism of Solid Sequencing

1. Template Preparation

   • Single-stranded DNA (ssDNA) is immobilized on a solid support.

2. Probe Hybridization

   • Short, fluorescently labeled oligonucleotide probes are hybridized to the template DNA.

3. Ligation by DNA Ligase

   • DNA ligase joins the correctly matched probes to the template strand.

4. Detection

   • The sequence is read based on the fluorescent signals emitted by the probes.

5. Cleavage and Repetition

   • The ligated probes are removed, and the process is repeated for the next round of sequencing.

Role of DNA Ligase in Solid Sequencing

1. Joining Probes

DNA ligase catalyzes the formation of phosphodiester bonds between adjacent nucleotides, ensuring that the correct probes are joined to the template.

2. Increasing Sequencing Accuracy

The specificity of DNA ligase in recognizing correct base pairing improves the accuracy of sequencing.

3. High Throughput

Solid sequencing technology allows the parallel processing of millions of sequences, making it suitable for large-scale genomics studies.

Why DNA Ligase is Essential in Solid Sequencing

• DNA ligase enables highly accurate sequence determination by ligating probes only when a perfect match occurs.
• It ensures that the hybridization is specific, reducing sequencing errors.
• The ability to use short probes increases the sensitivity and resolution of sequencing.

Advantages of Solid Sequencing Technology

1. High Accuracy

Solid sequencing has a low error rate due to the specificity of DNA ligase.

2. Multiplexing Capability

Multiple sequences can be processed simultaneously, increasing throughput.

3. Short Read Lengths

Short oligonucleotide probes improve resolution and enable detailed mapping of genomic regions.

Challenges in Solid Sequencing Technology

Comparison of Solid Sequencing with Other NGS Techniques

Applications of Solid Sequencing Technology

✅ 1. Whole Genome Sequencing

Used for sequencing entire genomes with high accuracy.

✅ 2. Transcriptome Analysis

Enables the identification of gene expression patterns.

✅ 3. Cancer Genomics

Detects mutations and structural variations associated with cancer.

✅ 4. Microbial Genomics

Used to sequence microbial genomes and identify antibiotic resistance genes.

Importance for CSIR NET and Other Competitive Exams

Solid sequencing is a frequently asked topic in exams like CSIR NET Life Science, IIT JAM, GATE Biotechnology, and DBT JRF. Understanding its mechanism, advantages, and enzyme usage is essential for scoring high in molecular biology sections.

Conclusion

Solid sequencing technology is a powerful and accurate method for genome sequencing. The use of DNA ligase enhances the specificity and accuracy of the process, making it suitable for complex genomic studies. Mastering the principles of solid sequencing is essential for students preparing for CSIR NET, IIT JAM, GATE Biotechnology, and DBT JRF.

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FAQs

Q1. Why is DNA ligase used in solid sequencing?
DNA ligase ensures accurate joining of probes during sequencing, improving accuracy.

Q2. What is the main advantage of solid sequencing over Sanger sequencing?
Solid sequencing allows high-throughput and parallel sequencing, increasing efficiency.

Q3. How does solid sequencing differ from Illumina sequencing?
Solid sequencing uses DNA ligase, while Illumina sequencing relies on DNA polymerase.

This article was prepared with insights from Let’s Talk Academy, the top coaching institute for competitive life science exams.