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However, mate-pair approaches have the added advantage of providing paired sequence from the ends of comparatively large DNA fragment sizes, which promotes the recovery of reads that span insertion sites.
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The amplified fragments spanned insertion termini.
However, solely depending on automated methods and software can leave many reads spanning insertions and deletions (indels) misaligned; therefore postreviewing the data for mismapping is always a good practice, especially in the candidate regions.
As single base pair insertion in CrWRKY8 was not clear by aligning other copies of this contig, the region spanning the insertion was cloned for verification.
RT-PCR performed using primers which span an insertion site may not be sufficient to conclude that all activity of a protein of interest is abolished.
RT-PCR experiments, with primers that span the insertion sites, were neither able to detect any HAM1 mRNA in the ham1 mutants nor HAM2 mRNA in ham2 mutant (Fig. 2B).
P element excision is imprecise and deletions of varying size spanning the insertion site or extending from the insertion site in either direction are often recovered at significant frequencies [43].
A 1.6-Mb region that spans this insertion site (rux to cm ) is five map units, for an average of 3.13 map units per Mb.
This situation can arise, for example, when the junction sequence spans an insertion or deletion of several bases in the sample relative to the reference.
This problem was probably caused by the presence of "hybrid" reads including sequences of two different elements (spanning their insertion sites or recombination breakpoints, see also Discussion).
In particular, when a contig/read spans an insertion site around a breakpoint that is farther from the duplication original site (Fig. 4A), then the alignment procedure is exactly the same as for sequences with simple deletion or insertion.
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