Sequencing Techniques Quiz

This technique uses dideoxynucleotides (ddNTPs) to terminate DNA chain elongation during sequencing. Each ddNTP lacks a 3' hydroxyl group, preventing further nucleotide addition. The resulting DNA fragments of varying lengths are then separated by electrophoresis, revealing the sequence.

Pyrosequencing is a high-throughput method that measures the release of pyrophosphate (PPi) during DNA synthesis. It involves a series of enzymatic reactions that convert PPi into light, which is detected to determine the nucleotide incorporated. This process is faster and more efficient than traditional sequencing methods, allowing for large-scale sequencing projects.

This approach involves walking along a chromosome by repeatedly cloning and sequencing overlapping DNA fragments. By creating contiguous sequences, the complete sequence of a chromosome can be assembled. It is a powerful technique for mapping large genomes.

SOLiD sequencing relies on the ligation of short, fluorescently labeled oligonucleotide probes to a DNA template. Each probe corresponds to a specific nucleotide sequence. By detecting the colors of the ligated probes, the DNA sequence can be determined. This method is known for its high accuracy.

Massively parallel pyrosequencing involves simultaneously sequencing millions of DNA fragments using a microfluidic chip. Each fragment is attached to a bead, and the pyrosequencing reaction proceeds on each bead individually. This parallel processing enables rapid and high-throughput sequencing, producing huge amounts of data.