ngsfilter
Hello!
I'm having problems in demultiplexing my sequences with ngsfilter. Essentially, the program assigns sequences just to the sample belonging to the first raw of my sample_description.txt file, ignoring all the other tag combinations below, while in the "unassigned" file I found various existing tag combinations. Furthermore, in most of the assigned sequences, ngsfilter is not able to assign the forward tag, the reverse tag or both tags to sequences. Here I attach two sequences taken from the output of my ngsfilter run.
K00253:122:HFGGKBBXY:7:1101:16691:1226_CONS_SUB_CMP ali_length=91; seq_ab_match=91; tail_quality=36.5; reverse_match=gggtatctaatcccagtttg; seq_a_deletion=0; reverse_score=80.0; reverse_primer=gggtatctaatcccagtttg; merged_sample={'b01_1': 1}; seq_a_mismatch=0; seq_b_mismatch=0; score=363.844489406; mid_quality=61.2827225131; avg_quality=58.9336492891; seq_a_single=60; score_norm=3.99829109237; status=partial; direction=reverse; seq_b_insertion=0; experiment=1; seq_b_deletion=0; seq_a_insertion=0; seq_length_ori=211; count=1; seq_length=181; mode=alignment; head_quality=36.5; seq_b_single=60; ntcgtgccagccaccgcggttatacgagaggcccaagttgataggtaacggcgtaaaggg tggttagggagtactatacagtaaagccgaacgtctatcaatgttgtttgaagctctcga agattggaagccccgccacgaaagtgactttaccatccctgaatccacgaaagccagggc a K00253:122:HFGGKBBXY:7:1101:6492:1244_CONS_SUB_SUB_CMP status=full; seq_a_deletion=0; forward_primer=aaactcgtgccagccacc; reverse_primer=gggtatctaatcccagtttg; merged_sample={'b01_1': 1626}; experiment=1; seq_b_insertion=0; seq_b_deletion=0; seq_a_insertion=0; reverse_tag=acacacac; count=1626; seq_length=163; mode=alignment; gcggttatacgaggggcccaagttgacagaagacggcgtaaagcgtggttaatgagagat acaataaagccaaattccatcaaggctgtcatacgcatccgatgacgagaggaaccccag cgaaagtgactttagtaagcatgaggccacgaaagctagggaa
As you can see, both sequences are assigned to the sample b01_1, which corresponds to the sample on the first raw of the sample_description.txt file. TELE02_sample_description.txt. I may have mistaken something in the preceding steps, or in the text file format. Does anybody have the same issue or any suggestion? Thank you very much!