| Genomic DNA extraction |
Qiagen DNeasy |
Qiagen DNeasy |
Salting out |
| NGS Technology |
IonTorrent | Ion318Chip |
IonTorrent | Ion318Chip |
Illumina® MiSeq |
| Library preparation |
Iron ExpressTM |
Iron ExpressTM |
|
| Type of reads |
Single reads |
Single reads |
Paired-end reads |
| Number of reads |
1,374,891 |
1,587,789 |
2,800,640 |
| Merged reads |
– |
– |
1,169,198 |
| Reads length |
8–620 |
8–618 |
300 |
| Merged reads length |
– |
– |
300–575 |
| FASTQC |
|
|
|
| % GC |
41 |
42 |
39 |
| Sequence quality < Phred 20 |
yes at positions >325 bp |
yes at positions >350 bp |
no |
| Per sequence quality—Phred score |
29 |
29 |
36 |
| Sequence length distribution |
peak at 350 bp |
peak at 350 bp |
plateau at 520–540 bp |
| QDD2 |
|
|
|
| QDD2 pipe 1—Sequence preparation and microsatellite detection |
| Number sequence length ≥80 bp |
1,235,685 |
1,405,082 |
1,169,198 |
| Number sequence with microsatellite |
109,641 (8.9%) |
167,702 (11.9%) |
130,269 (11.1%) |
| QDD2 pipe 2—Sequence similarity detection |
| Total # input sequences |
109,641 |
167,702 |
130,269 |
| Numer of unique consensus sequences |
18,978 |
N/A |
N/A |
| Number of singleton sequences |
49,122 |
N/A |
N/A |
| Number of reads in output |
68,100 |
63,789 |
69,714 |
| QDD2 pipe 3—Primer design |
| Total number of input sequences |
68,100 |
63,789 |
69,714 |
| Total number of sequences with target MS |
67,461 |
63,785 |
69,714 |
| Total number of sequences with primers |
29,485 |
20,233 |
19,867 |
| Filtering QDD output |
|
|
|
| Total # input sequences |
29,485 |
20,233 |
19,867 |
| Total # sequences after filtering criteria |
121 |
97 |
103 |