Table 1 Datasets generated in this study and the results of barcoding using ONTbarcoder at 200X coverage (Consensus by Length) and 100X coverage (Consensus by Similarity)
Dataset name | Flow cell details Run time/Guppy version | Raw reads/reads passing length threshold/reads of suitable length/ demultiplexed | Demultiplexing rate/# QC_compliant barcodes/# Filtered barcodes with 1 N/# Filtered barcodes with > 1 N /# Unreliable barcodes |
|---|---|---|---|
MinION R10.3 datasets | |||
Mixed Diptera (658 bp, N=511) | R10.3: reused flow cell: 71 pores according to QC, but 500+ active during run Runtime: 27.5 h Guppy: 4.2.3+f90bd04 | 3,864,000/3,425,357/3,560,389/1,544,758 | 43.39%/495/2/5/8 Total success rate= 502/511 (98.2%) |
Afrotropical Phoridae (658 bp, N=4275) | R10.3: new flow cell: QC: 1101 pores Runtime: 49.5 h Guppy: 4.0.11+f1071ce | 6,838,903/5,465,164/5,474,306/2,681,029 | 48.97%/3722/121/59/247 Total success rate= 3905/4275 (91.3%) |
Palaearctic Phoridae (658 bp, N=9932) | R10.3: new flow cell: QC: 1239 pores Runtime: 47.5 h Guppy: 4.2.3+f90bd04 | 16,595,984/15,658,174/16,100,505/5,012,489 | 31.13%/8026/108/231/780 Total success rate= 8365/9932 (84.2%) |
Palaearctic Phoridae (313 bp, N=9929) | R10.3: new flow cell: QC: 1297 pores Runtime: 37 h Guppy: 4.2.3+f90bd04 | 13,690,869/13,221,764/10,366,455/12,983,260/2,015,135 | 15.52%/8705/118/112/899 Total success rate= 8935/9929 (90%) |
Flongle datasets | |||
Mixed Diptera Subsample (658 bp, N=257) | Flongle: new QC: 81 pores Runtime: 24 h Guppy: v 4.0.11+f1071ce | 294,896/222,189/190,952/33,270 | 17.42%/185/35/20/9 Total success rate= 240/257 (93.4%) |
Chironomidae (313 bp, N=191) | Flongle: new QC: 74 pores Runtime: 15 h Guppy: 4.2.3+f90bd04 | 560,062/525,087/504,621/108,574 | 21.52%/178/1/2/6 Total success rate= 181/191 (94.8%) |