Lecture 11 molecular genetics

• What type of reaction is Sanger Sequencing?

  DNA polymerase reaction

  
• What components are required for Sanger Sequencing?

  DNA template, primers, dNTPs, polymerase, Mg2+

  
• What is the role of dideoxynucleotide triphosphates in Sanger Sequencing?

  They randomly stop the polymerase reaction

  
• What do dideoxynucleotides lack?

  A 3’-OH group
• What is the purpose of using fluorophores in Sanger Sequencing?

  To detect specific colors for bases

  
• What does a chromatogram represent in Sanger Sequencing?

  Results of the sequencing reactions

  
• What are the key features of high-throughput sequencing?

  • Many reactions in parallel
  • Utilizes microfluidic technologies
  • Outputs several Gb to Tb per run
• What is the main advantage of high-throughput sequencing over Sanger Sequencing?

  Cost per base pair is much cheaper
• Why is Sanger Sequencing still useful despite high-throughput methods?

  For one or a small number of sequences

  
• What is a challenge associated with high-throughput sequencing data?

  Bioinformatic challenges in analyzing data
• What is a hybrid assembly in sequence analysis?

  Combining short and long DNA reads
• What are the steps in sequence analysis for alignment?

  1. Align multiple samples for the same region
  2. Look for SNPs or other differences
• What is the purpose of creating phylogenetic trees in sequence analysis?

  • Show relationships between genes/DNA regions
  • Determine if genes are homologous
• How do nanopore technologies differ from traditional sequencing methods?

  They move DNA through pores to detect bases

  
• What is a potential application of nanopore sequencing technology?

  Real-time field sequencing
• What is the output time range for high-throughput sequencing runs?

  24 hours to several days
• Which companies are known for nanopore sequencing technologies?

  Oxford Nanopore Technologies and PacBio