Table 3 Comparison of barcodes obtained by ONTbarcoder with NGSpeciesID (green=highest number of correct and lowest number of errors for datasets). NGSpeciesID was applied once to the demultiplexed reads obtained with ONTbarcoder and once to those obtained with minibar. Barcode calling was done using all demultiplexed reads as well as 200X subset. ONTbarcoder was run at 200X read coverage (Consensus by Length) and 100X coverage (Consensus by Similarity). The accuracy of MinION barcodes is only compared to reference barcodes obtained with Illumina/Sanger sequencing. Errors are defined as the sum of substitution and indel errors
ONTBarcoder 200X | ONTBarcoder dem1 + NGSpeciesID All reads | ONTBarcoder dem1 + NGSpeciesID 200X | Minibar + NGSpeciesID All reads | Minibar + NGSpeciesID 200X | |
|---|---|---|---|---|---|
Mixed Diptera Subsample Flongle | |||||
Identical+compatible2(# of reference barcodes) | 226 (231) | 72 (241) | 73 (241) | 123 (241) | 122 (241) |
Identical* | 177 | 70 | 71 | 121 | 120 |
Incorrect, (1/2/3/4/5+) errors | 5 (3/1/1/0/0) | 169 (83/60/13/6/7) | 168 (88/55/13/7/5) | 118 (29/21/8/4/56) | 119 (31/20/8/4/56) |
# reads demultiplexed/reads of length 658±50 | 33,270/32,978 | 19,511/15,935 | |||
Mixed Diptera R10.3 | |||||
Identical+compatible2(# of reference barcodes) | 474 (476) | 173 (478) | 353 (478) | 418 (478) | 453 (478) |
Identical* | 466 | 170 | 347 | 413 | 447 |
Incorrect, with (1/2/3/4/5+) errors | 2 (0/0/1/0/1) | 305 (126/97/48/17/17) | 125 (91/24/5/1/4) | 60 (45/9/3/0/3) | 25 (20/2/0/0/3) |
# reads demultiplexed/reads of length 658±50 | 1,544,768/1,532,408 | 1,540,751/1,378,910 | |||
Afrotropical Phoridae R10.3 | |||||
Identical+compatible2 (# of reference barcodes) | 3293 (3321) | 2832 (3339) | 3111 (3341) | 3193 (3340) | 3245 (3340) |
Identical | 3239 | 2832 | 3111 | 3193 | 3245 |
Incorrect, (1/2/3/4/5+) errors | 28 (15/5/0/0/8) | 507 (362/90/19/12/24) | 230 (172/31/5/3/19) | 147 (90/27/3/1/26) | 95 (50/13/12/1/27) |
# reads demultiplexed/reads of length 658±50 | 2,681,029/2,668,933 | 3,117,597/2,782,367 | |||