An Efficient DNA Sequencing Strategy Based on the Bacteriophage Mu in Vitro DNA Transposition Reaction

Table 3.

Comparison of the Characteristics of Mu and Ty1 Sequencing Strategies

Mu Ty1
Reaction catalyzed by MuA transposase Ty1 virus-like particles
Donor DNA length (kb) 1.2 0.9
Introduction of reaction products into cells transformation/electroporation electroporation
Reaction efficiency
 maximum reported (insertions per target) 1/10 1/2.5 × 103
Target site duplication (bp) 5 5
Standard reaction
 donor used (ng) 18 100–500
 target used (ng) 500 1000
 molar ratio of transposon to target 1:2 10:1
Reported recovery of clones
 minipreps studied (no., %) 95, 100 153, 100
 double insertions/cotransformants (%) 2 8
 unknown plasmid map (%) 0 3
 no transposon (%) 0 <1
 single insertion in one plasmid (%) 98 88
  • Mu characteristics (this study); Ty1 characteristics (Devine and Boeke 1994).

  • Allet (1979); Kahmann and Kamp (1979); Mizuuchi and Mizuuchi (1993); this study.

  • Eichinger and Boeke (1988, 1990); Ji et al. (1993);Braiterman and Boeke (1994); Primer Island Transposition Kit Protocol (Applied Biosystems).

  • The numbers for the Ty1 system were calculated from Table 2 of Devine and Boeke (1994). For comparison the number of minipreps studied was adjusted to 100%

This Article

  1. Genome Res. 9: 308-315

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