Figure
17. Sample Prep Workflow.
The
input sample is first fragmented to the desired size. The ends are repaired
and the hairpin structures are ligated to the ends of each
fragment.
A size selection and purification step selects those fragments with the
adaptors attached to both ends. The SMRTbell templates
then
can go through the sequencing reaction. A strand displacing polymerase enzyme
opens the SMRTbell into a circular template and can
generate
independent reads, both forward and reverse of the same DNA molecule. The
quality score increases linearly with the number of
times
the molecule is sequenced.