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Second-generation DNA Sequencing Technique-Roche 454: Pryrosequencing,…

- - - - Sequencing by ligation (SBL) or sequencing by synthesis (SBS) (Kchouk et al., 2017)
      - High-throughput DNA sequencing
      - High sensitivity
      - Faster sequencing than in the first generation and low cost
      - Sample preparation required PCR amplification (Kchouk et al., 2017)
      - Shorter read length than in the Sanger sequencing
      - The sequencing result is detected directly without the requirement of electrophoresis (Kchouk et al., 2017)
      - Adapted to a wide range of cases
  - - - Physical inspection of DNA molecules using a nanopore
      - Single-molecule real-time sequencing approach (SMRT)
      - DNA sequencing at higher throughput
      - High sensitivity
      - Time required much less than SGS and lower cost
      - Simple sample preparation without PCR amplification
  - - - Uses Chain termination method
      - Uses one of the double-stranded DNA strands as a template for sequencing (Kchouk et al., 2017)
      - Low-throughput DNA sequencing
      - Low sensitivity
      - Time-consuming and high cost
      - Low radioactivity (Kchouk et al., 2017)
      - Preparation of the sample requires PCR amplification and recombinant DNA
      - Long read length
      - Suitable for small number of cases
- - - - STEP 2: Loading DNA Sample onto Beads
      - STEP 1: Sample Preparation
      - STEP 3: Sequencing
  - - - Data generated by 454 Sequencing™ on the Genome Sequencer FLX
      - Analysis of the genome.
      - Multitude of 454 Sequencing Flowgrams
      - Millions of DNA Sequences
      - Reference DNA Sequence
      - Assembled Human Genome Sequence
- - - - -downstream bioinformatics resulting in sequence assembly with longer contigs
      - higher N50 length
      - less gaps specially in de novo sequencing projects