Fluorescent dye terminator sequencing

Definition

Fluorescent dye terminator sequencing is a method of DNA sequencing that uses fluorescently labeled dideoxynucleotides to terminate DNA strand elongation at specific bases. This technique allows for the simultaneous detection of multiple fluorescent dyes, enabling the identification of the DNA sequence in a highly efficient manner. By incorporating different dyes for each of the four nucleotides, this method enhances the accuracy and speed of sequencing compared to traditional methods.

5 Must Know Facts For Your Next Test

1. Fluorescent dye terminator sequencing improves upon Sanger sequencing by using multiple fluorescent dyes to label the dideoxynucleotides, allowing for the detection of all four bases in a single reaction.
2. The method typically utilizes a capillary electrophoresis system, where DNA fragments are separated by size, and a laser detects the fluorescent signals emitted from each fragment.
3. This sequencing technique can generate high-throughput results, making it suitable for large-scale genomic projects and applications such as whole genome sequencing.
4. Fluorescent dye terminator sequencing has a lower error rate compared to older methods, providing higher accuracy in determining nucleotide sequences.
5. The use of automated sequencers has made fluorescent dye terminator sequencing more accessible, enabling rapid and cost-effective DNA analysis for various research and clinical applications.

Review Questions

• How does fluorescent dye terminator sequencing improve upon traditional Sanger sequencing methods?
  • Fluorescent dye terminator sequencing enhances traditional Sanger sequencing by incorporating different fluorescent dyes for each of the four dideoxynucleotides. This allows for simultaneous detection of all four bases during a single reaction, significantly increasing throughput and efficiency. The use of automated systems further accelerates the process, reducing the time needed to obtain sequencing results while improving accuracy compared to earlier methods.
• Discuss the role of capillary electrophoresis in the process of fluorescent dye terminator sequencing and how it contributes to data analysis.
  • Capillary electrophoresis plays a crucial role in fluorescent dye terminator sequencing by separating DNA fragments based on their size as they migrate through a capillary filled with a gel matrix. When a laser excites the fluorescent labels on the fragments, it generates signals corresponding to each base, which are detected and recorded. This technique allows for high-resolution separation and accurate reading of the sequence data, enabling reliable analysis and interpretation of the results.
• Evaluate the significance of fluorescent dye terminator sequencing in the context of modern genomics and its impact on genetic research.
  • Fluorescent dye terminator sequencing has revolutionized modern genomics by providing an efficient and high-throughput method for determining DNA sequences. Its impact is profound, as it enables large-scale projects like whole genome sequencing and personalized medicine initiatives. The lower error rates associated with this method enhance confidence in genetic research findings, facilitating advances in understanding genetic disorders, evolution, and population genetics. Ultimately, this technology continues to shape our approach to genomic studies and biological discoveries.

Related terms

Dideoxynucleotide (ddNTP): A modified nucleotide lacking a hydroxyl group on the 3' carbon, which prevents further extension of the DNA strand when incorporated.

Sanger sequencing:

A widely used method of DNA sequencing based on selective incorporation of chain-terminating dideoxynucleotides during DNA replication.

Capillary electrophoresis:

A technique used to separate the fragments generated in fluorescent dye terminator sequencing based on their size and charge, allowing for the detection of each labeled DNA fragment.