Teichlab / bracer

BraCeR - reconstruction of B cell receptor sequences from single-cell RNAseq data
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test results differ from expected #60

Closed zhouxinseeu closed 3 years ago

zhouxinseeu commented 3 years ago

Hi! I have run bracer test and have different results, but there is no error when testing. Is it common? my results:

BCR_H reconstruction:   2 / 3 (66.7%)
BCR_K reconstruction:   0 / 3 (0.0%)
BCR_L reconstruction:   3 / 3 (100.0%)

Paired HK productive reconstruction:    0 / 3 (0.0%)
Paired HL productive reconstruction:    2 / 3 (66.7%)
Paired KL productive reconstruction:    0 / 3 (0.0%)

+--------+----------------+---------------+----------------+
|        | 0 recombinants | 1 recombinant | 2 recombinants |
+--------+----------------+---------------+----------------+
| all H  | 1              | 2 (100%)      | 0 (0%)         |
| all K  | 2              | 1 (100%)      | 0 (0%)         |
| all L  | 0              | 3 (100%)      | 0 (0%)         |
| prod H | 1              | 2 (100%)      | 0 (0%)         |
| prod K | 3              | 0 (N/A%)      | 0 (N/A%)       |
| prod L | 0              | 3 (100%)      | 0 (0%)         |
+--------+----------------+---------------+----------------+

##Cells with more than two recombinants for a locus##

The following cells are likely multiplets or contaminated as they contain more than two recombinants for a locus. Consider removing them from downstream analysis.
None

##Proportion of full-length sequences of all recovered sequences##

        H       K       L
all     2/2 (100%)      1/1 (100%)      3/3 (100%)
prod    2/2 (100%)      0/0 (N/A%)      3/3 (100%)

##Isotype of cells with productive heavy chain##

Isotype cells   % of cells
IGHA1   2       100.00

#Clonotype groups#
This is a text representation of the groups shown in clonotype_network_with_identifiers.pdf.
It does not exclude cells that only share heavy chain and not light chain if Summarise is run with --IGH_networks.

cell2, cell3

#Cells with no reconstructed sequences#
None

expected results:

BCR_H reconstruction:   3 / 3 (100.0%)
BCR_K reconstruction:   0 / 3 (0.0%)
BCR_L reconstruction:   3 / 3 (100.0%)

Paired HK productive reconstruction:    0 / 3 (0.0%)
Paired HL productive reconstruction:    3 / 3 (100.0%)
Paired KL productive reconstruction:    0 / 3 (0.0%)

+--------+----------------+---------------+----------------+
|        | 0 recombinants | 1 recombinant | 2 recombinants |
+--------+----------------+---------------+----------------+
| all H  | 0              | 3 (100%)      | 0 (0%)         |
| all K  | 2              | 1 (100%)      | 0 (0%)         |
| all L  | 0              | 3 (100%)      | 0 (0%)         |
| prod H | 0              | 3 (100%)      | 0 (0%)         |
| prod K | 3              | 0 (N/A%)      | 0 (N/A%)       |
| prod L | 0              | 3 (100%)      | 0 (0%)         |
+--------+----------------+---------------+----------------+

##Cells with more than two recombinants for a locus##

The following cells are likely multiplets or contaminated as they contain more than two recombinants for a locus. Consider removing them from downstream analysis.
None

##Proportion of full-length sequences of all recovered sequences##

        H       K       L
all     3/3 (100%)      1/1 (100%)      2/3 (67%)
prod    3/3 (100%)      0/0 (N/A%)      2/3 (67%)

##Isotype of cells with productive heavy chain##

Isotype cells   % of cells
IGHA1   2       66.67
IGHG1   1       33.33

#Clonotype groups#
This is a text representation of the groups shown in clonotype_network_with_identifiers.pdf.
It does not exclude cells that only share heavy chain and not light chain if Summarise is run with --IGH_networks.

cell2, cell3

#Cells with no reconstructed sequences#
None

test log:

56438 reads; of these:
  56438 (100.00%) were paired; of these:
    35350 (62.64%) aligned concordantly 0 times
    21088 (37.36%) aligned concordantly exactly 1 time
    0 (0.00%) aligned concordantly >1 times
    ----
    35350 pairs aligned concordantly 0 times; of these:
      73 (0.21%) aligned discordantly 1 time
    ----
    35277 pairs aligned 0 times concordantly or discordantly; of these:
      70554 mates make up the pairs; of these:
        69268 (98.18%) aligned 0 times
        232 (0.33%) aligned exactly 1 time
        1054 (1.49%) aligned >1 times
38.63% overall alignment rate
56438 reads; of these:
  56438 (100.00%) were paired; of these:
    56438 (100.00%) aligned concordantly 0 times
    0 (0.00%) aligned concordantly exactly 1 time
    0 (0.00%) aligned concordantly >1 times
    ----
    56438 pairs aligned concordantly 0 times; of these:
      0 (0.00%) aligned discordantly 1 time
    ----
    56438 pairs aligned 0 times concordantly or discordantly; of these:
      112876 mates make up the pairs; of these:
        112876 (100.00%) aligned 0 times
        0 (0.00%) aligned exactly 1 time
        0 (0.00%) aligned >1 times
0.00% overall alignment rate
56438 reads; of these:
  56438 (100.00%) were paired; of these:
    48319 (85.61%) aligned concordantly 0 times
    8119 (14.39%) aligned concordantly exactly 1 time
    0 (0.00%) aligned concordantly >1 times
    ----
    48319 pairs aligned concordantly 0 times; of these:
      114 (0.24%) aligned discordantly 1 time
    ----
    48205 pairs aligned 0 times concordantly or discordantly; of these:
      96410 mates make up the pairs; of these:
        95663 (99.23%) aligned 0 times
        195 (0.20%) aligned exactly 1 time
        552 (0.57%) aligned >1 times
15.25% overall alignment rate
56438 reads; of these:
  56438 (100.00%) were paired; of these:
    31766 (56.28%) aligned concordantly 0 times
    24672 (43.72%) aligned concordantly exactly 1 time
    0 (0.00%) aligned concordantly >1 times
    ----
    31766 pairs aligned concordantly 0 times; of these:
      270 (0.85%) aligned discordantly 1 time
    ----
    31496 pairs aligned 0 times concordantly or discordantly; of these:
      62992 mates make up the pairs; of these:
        61549 (97.71%) aligned 0 times
        144 (0.23%) aligned exactly 1 time
        1299 (2.06%) aligned >1 times
45.47% overall alignment rate

[build] loading fasta file /Personal/zhouxin/software/bracer/test_data/results/cell1/expression_quantification/kallisto_index/cell1_transcriptome.fa
[build] k-mer length: 31
[build] warning: clipped off poly-A tail (longer than 10)
        from 1827 target sequences
[build] warning: replaced 4 non-ACGUT characters in the input sequence
        with pseudorandom nucleotides
[build] counting k-mers ... done.
[build] building target de Bruijn graph ...  done 
[build] creating equivalence classes ...  done
[build] target de Bruijn graph has 1510231 contigs and contains 142452662 k-mers 

[quant] fragment length distribution will be estimated from the data
[index] k-mer length: 31
[index] number of targets: 231,959
[index] number of k-mers: 142,452,662
[index] number of equivalence classes: 990,749
[quant] running in paired-end mode
[quant] will process pair 1: /Personal/zhouxin/software/bracer/test_data/cell1_1.fastq
                             /Personal/zhouxin/software/bracer/test_data/cell1_2.fastq
[quant] finding pseudoalignments for the reads ... done
[quant] processed 56,438 reads, 41,888 reads pseudoaligned
[quant] estimated average fragment length: 135.93
[   em] quantifying the abundances ... done
[   em] the Expectation-Maximization algorithm ran for 246 rounds

/SGRNJ/Public/Software/conda_env/Bracer/lib/python3.7/site-packages/seaborn/distributions.py:2557: FutureWarning: `distplot` is a deprecated function and will be removed in a future version. Please adapt your code to use either `displot` (a figure-level function with similar flexibility) or `histplot` (an axes-level function for histograms).
  warnings.warn(msg, FutureWarning)
/SGRNJ/Public/Software/conda_env/Bracer/lib/python3.7/site-packages/seaborn/distributions.py:306: UserWarning: Dataset has 0 variance; skipping density estimate.
  warnings.warn(msg, UserWarning)
/SGRNJ/Public/Software/conda_env/Bracer/lib/python3.7/site-packages/seaborn/distributions.py:2557: FutureWarning: `distplot` is a deprecated function and will be removed in a future version. Please adapt your code to use either `displot` (a figure-level function with similar flexibility) or `histplot` (an axes-level function for histograms).
  warnings.warn(msg, FutureWarning)

thank you!