DeepBinner on SQK-RLB001 - Rapid Low Input by PCR Barcoding Kit
Opened this issue · 0 comments
Dear Ryan, thanks a lot for all your very useful tools for Nanopore data handling.
I'm currently trying to demultiplex a MinION run (FLO-MIN106) barcoded with SQK-RLB001 - Rapid Low Input by PCR Barcoding Kit.
Barcodes 03; 04; 05; 06; 07; 08; 09 were used (tot. 7 barcodes)
I gave a shot at deepBinner using the Rapid model and while a lot of reads are not getting classified, there is still a fair amount that got assigned a barcode using default threshold. I was wondering if those results can be trusted in anyway ? My strategy would have been to then basecall each fast5 barcode directories with demultiplexing and then concatenate fasta for which both methods agrees, as you're suggesting for maximum stringency.
As I'm assembling fosmid inserts, I'm not too much scared by read yield but very much by barcode misclassifications.
Barcode Count
1 45
2 134
3 5639
4 1819
5 889
6 1754
7 1042
8 1782
9 1060
10 574
11 494
12 170
none 10198
Found 66,997 fast5 files in Fast5_20180131_1523_7fosmid_RLB001-pass/
Classifying fast5s: 25600 / 25600 (100.0%)
Moving fast5s: 25600 / 25600 (100.0%)
Barcode Count
1 33
2 98
3 5818
4 2010
5 1076
6 1985
7 1091
8 1964
9 1074
10 364
11 388
12 94
none 9605
Found 80,557 fast5 files in Fast5_20180131_1523_7fosmid_RLB001-pass/
Classifying fast5s: 25600 / 25600 (100.0%)
Moving fast5s: 25600 / 25600 (100.0%)
Barcode Count
1 42
2 111
3 5898
4 2041
5 1073
6 1963
7 1017
8 1984
9 1110
10 278
11 391
12 99
none 9593