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Figure 4 | Genome Biology

Figure 4

From: Rapid, low-input, low-bias construction of shotgun fragment libraries by high-density in vitro transposition

Figure 4

Insert size showing steric hindrance. (a) Insert size was generated from libraries spiked into a paired-end 101 bp run resulting in a large proportion of reads reading into the adaptor sequence. Tails of reads were then aligned to one another to discern the insert size between adaptors, resulting in a mapping-independent insert size at the lower extreme. All reads with an insert size less than 25 bp were PCR artifacts. (b) The noticeable drop below 40 bp is consistent with a model for complete saturation of transposition events on a given stretch of DNA. The roughly 110 Å transposase homodimer (grey) is bound to genomic DNA (blue), such that the core of the enzyme acts on a 9 bp region drawn out to 41 Å as well as approximately 10 additional bases of DNA flanking either side (~34 Å each) that are essentially protected from a subsequent transposase attack due to steric hindrance. Since the core region is duplicated in the process, the minimum spacing of transposition events is approximately 38 bp.

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