Long read sequencing technology to solve complex genomic regions assembly in plants
Résumé
Background: Numerous completed or on-going whole genome sequencing projects have highlighted the fact
that obtaining a high quality genome sequence is necessary to address comparative genomics questions such as structural variations among genotypes and gain or loss of specific function. Despite the spectacular progress thathas been made in sequencing technologies, obtaining accurate and reliable data is still a challenge, both at the whole genome scale and when targeting specific genomic regions. These problems are even more noticeable for complex plant genomes. Most plant genomes are known to be particularly challenging due to their size, high density of repetitive elements and various levels of ploidy. To overcome these problems, we have developed a strategy to reduce genome complexity by using the large insert BAC libraries combined with next generation sequencing technologies.
Results: We compared two different technologies (Roche-454 and Pacific Biosciences PacBio RS II) to
sequence pools of BAC clones in order to obtain the best quality sequence. We targeted nine BAC clones from
different species (maize, wheat, strawberry, barley, sugarcane and sunflower) known to be complex in terms of
sequence assembly. We sequenced the pools of the nine BAC clones with both technologies. We compared
assembly results and highlighted differences due to the sequencing technologies used.
Conclusions: We demonstrated that the long reads obtained with the PacBio RS II technology serve to obtain
a better and more reliable assembly, notably by preventing errors due to duplicated or repetitive sequences in the
same region.
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