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Table 1 Comparison of ABI-Sanger and 454 sequencing procedures

From: 454 sequencing put to the test using the complex genome of barley

  ABI-Sanger 454 time requireda
Isolation of BAC DNA x x 1 day
Mechanical shearing x x 2 h
Cloning x   4 h
Clone picking x   2 h
Plasmid DNA extraction x   20 h
Reactions on thermocycler x   36 h
Clean up reaction products x   2 h
ABI 3730xl sequencer run x   24 h
454 sequencing library   x 4 hb
Amplification in PCR microreactors   x 6 hb
GS 20 sequencing run   x 4 hb,c
Assembly of raw sequences x x days to weeks
  1. aThe procedures and estimated time requirements describe the process for sequencing a BAC clone with a size of 100 kb. For ABI-Sanger, numbers are calculated to reach an approximately 10-fold coverage.
  2. baccording to [5]
  3. cOne 454 GS20 run produces ~20 Mb, approximately 10 times more than required for a sufficient coverage (20 ×) of a 100 kb BAC clone.