Table 1

Failure mode categories Failed wells were distributed into each category based on observational data taken during sequencing pipeline procedures and manual evaluation of electropherogram traces.

Failure Mode
Trace characteristic
No. of sequencing reactions
Percent of all failed wells (Q20 < 600)

Blocked capillary
Noisy or no data with a low signal intensity value (<100). Verified with capillary control results.
64
5.5
Low signal strength*
Noisy or no data with a low signal intensity value (<100) that is very close to or falls below the instruments detectable limit.
310
26.5
Mixed clone w/ vector sequence
Clean vector sequence followed by noisy data immediately after the cloning site.
137
11.7
Mixed clone, no vector sequence
Noisy data throughout the trace with sufficient signal intensity.
27
2.3
Low signal to noise ratio
Discernable sequence peaks with strong intensity background noise.
22
1.9
Excess Dye peaks
Large dye front usually followed by noisy data.
10
0.9
Hardstop
Abrupt end to good sequence.
2
0.2
Repetitive Sequence
Long stretch of repetitive DNA sequence that is followed by slippage in sequence or noisy data.
17
1.5
Homopolymer stretch
Long stretch of a single nucleotide followed by slippage in sequence or noisy data.
99
8.4
Poly A Tail
Stretch of Ts (template A) followed by slippage in sequence or noisy data.
484
41.3
Total

1172
100.0

* Preliminary failure mode, further broken down to final failure modes in Table 2.

Yang et al. BMC Genomics 2005 6:2   doi:10.1186/1471-2164-6-2