Exploring Next Generation Sequencing Techniques

The video tutorial provides an overview of Next Generation Sequencing (NGS) and Sanger Sequencing, explaining their processes, applications, and differences. It covers various NGS platforms, including pyrosequencing, sequencing by synthesis, sequencing by ligation, and ion semiconductor sequencing, highlighting their unique features and limitations. The tutorial also compares the coverage and applications of these technologies and introduces the NGS services offered by abm.

What is one of the key advantages of Next Generation Sequencing (NGS) over traditional methods?

• It can sequence thousands to millions of DNA molecules simultaneously.

• It can sequence a single DNA molecule.

• It is slower than traditional methods.

• It is less accurate than traditional methods.

What is the primary use of Sanger Sequencing today?

• For sequencing lipids.

• For routine sequencing applications and validating NGS data.

• For sequencing RNA molecules.

• For sequencing proteins.

What initiates the DNA synthesis reaction in Sanger Sequencing?

• A double-stranded DNA template.

• A single primer complementary to the template.

• A mixture of four di-deoxynucleotides.

• A fluorescent dye.

What is a common feature of all Next Generation Sequencing platforms?

• They do not require sample preparation.

• They require a library obtained by amplification or ligation with custom adapter sequences.

• They use manual electrophoresis.

• They do not generate raw data.

Which NGS method uses the release of pyrophosphate to monitor the sequencing reaction?

• Pyrosequencing

• Ion semiconductor sequencing

• Sequencing by ligation

• Sequencing by synthesis

What is a disadvantage of pyrosequencing?

• Inability to sequence RNA.

• Short read lengths.

• High reagent cost and high error rate over strings of 6 or more homopolymers.

• Low throughput.

Which NGS method is used by Illumina platforms?

• Pyrosequencing

• Sequencing by synthesis

• Ion semiconductor sequencing

• Sequencing by ligation

What is a major disadvantage of sequencing by ligation?

• Short sequencing reads.

• High error rate.

• High cost.

• Low throughput.

What does ion semiconductor sequencing detect to determine the sequence of a cluster?

• Binding of oligonucleotide probes.

• Release of hydrogen ions.

• Fluorescent signal.

• Release of pyrophosphate.

Which NGS method is most popular for generating hundreds of coverage per run?

• Ion semiconductor sequencing

• Sequencing by ligation

• Sequencing by synthesis

• Pyrosequencing