If this error rate applied to the sequencing of the entire human mitochondrial genome comprising about 16,570 base pairs, then the majority of complete mtDNA sequences in a database would carry one or more incorrect bases – which was typically attained by the earliest sequencing attempts of the past, but most recent clinical mtDNA studies do not fare better and sometimes much worse [3] [5].
In prior single cell genome sequencing attempts, Zhang et al. [18] recovered 66% of a genome of a cultured Prochlorococcus strain in 477 contigs, while Marcy et al. [19] recovered an unknown fraction of a genome of an uncultured representative of the TM7 phylum in 288 contigs, with up to 10% Leptotrichia contamination.
Several sequencing attempts failed.
However, the sequencing of the samples could not confirm a mutation despite several sequencing attempts.
Out of 3,264 total sequencing attempts (large exons required sequencing several overlapping PCR products), 3,076 successful sequences were achieved.
Embryos that consistently failed at one or more loci after two sequencing attempts were excluded from parental analysis.
However, * collections from rare species where sampling may be difficult will still be useful for initial whole-genome sequencing attempts.
From 18,976 sequencing attempts 9,110 sequences remained after quality, vector and adapter trimming, and removal of mitochondrial DNA sequences (see Materials and methods).
From the 27,513 total 3' sequencing attempts on the tentatively unique cDNAs represented in Table 1, a total of 22,088 sequences met the criteria of high quality sequence.
For comparison purposes, we also performed the same sequencing process on 22 pairs of human Dukes B sporadic colorectal tumors and their matching-normal tissue samples, and obtained 8,342 successful sequences from 9,360 total sequencing attempts.
