Table 1

Comparison of ABI-Sanger and 454 sequencing procedures

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


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.

baccording to [5]

cOne 454 GS20 run produces ~20 Mb, approximately 10 times more than required for a sufficient coverage (20 ×) of a 100 kb BAC clone.

Wicker et al. BMC Genomics 2006 7:275   doi:10.1186/1471-2164-7-275

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