Fig 1.
Basic stages of the Single Molecule Real-Time (SMRT) DNA sequencing method.
SMRTbell adaptors are ligated onto the ends of a blunt-ended PCR amplicon to facilitate continuous sequencing of both strands of the amplicon. The entire sequence generated may include multiple copies of the sense and anti-sense strands of the PCR amplicon in a single read known as the Continuous Long Read (CLR). The post-sequencing bioinformatic post-processes are able to break down the CLR into shorter sub-reads, which encompass the sequence of one strand of the amplicon. These sub-reads can then be compared and used to create a consensus sequence.
Table 1.
Depth of coverage achieved using SMRT sequencing.
Table 2.
A comparison of expected HLA types, as typed by Anthony Nolan, with those generated by the Single Molecule Real-Time (SMRT) DNA Sequencing method.
Table 3.
Extensions and corrections to known HLA alleles identified in PacBio results.
Table 4.
Anomalies observed in the PacBio SMRT consensus sequences as compared to the expected allele.
Table 5.
Homopolymer count in 38 PacBio sequences (total length: 130117 bp).