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Commit e026e9ec authored by Celine Mercier's avatar Celine Mercier
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Fixed the new alignpaired end to work after ngsfilter with the 9879847 

possible cases
parent 4ddd1a1c
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Showing with 151 additions and 83 deletions
python/obitools3/commands/alignpairedend.pyx
View file @ e026e9ec
......@@ -15,16 +15,12 @@ from obitools3.libalign._qsrassemble import QSolexaRightReverseAssemble
from obitools3.libalign._solexapairend import buildConsensus, buildJoinedSequence
from obitools3.dms.obiseq cimport Nuc_Seq
from obitools3.libalign.shifted_ali cimport Kmer_similarity, Ali_shifted
from obitools3.commands.ngsfilter import REVERSE_SEQ_COLUMN_NAME, REVERSE_QUALITY_COLUMN_NAME
import sys
import os
REVERSE_SEQ_COLUMN_NAME = b"REVERSE_SEQUENCE"
REVERSE_QUALITY_COLUMN_NAME = b"REVERSE_QUALITY"
__title__="Aligns paired-ended reads"
......@@ -106,8 +102,7 @@ def alignmentIterator(entries, aligner):
else:
seqF = Nuc_Seq.new_from_stored(entries[i])
seqR = Nuc_Seq(seqF.id, seqF[REVERSE_SEQ_COLUMN_NAME], quality=seqF[REVERSE_QUALITY_COLUMN_NAME])
seqF.pop(REVERSE_SEQ_COLUMN_NAME)
seqF.pop(REVERSE_QUALITY_COLUMN_NAME)
seqR.index = i
ali = aligner(seqF, seqR)
......@@ -196,7 +191,7 @@ def run(config):
reverse = entries[1]
aligner = Kmer_similarity(forward, view2=reverse, kmer_size=config['alignpairedend']['kmersize'])
else:
aligner = Kmer_similarity(entries, kmer_size=config['alignpairedend']['kmersize'])
aligner = Kmer_similarity(entries, column2=entries[REVERSE_SEQ_COLUMN_NAME], qual_column2=entries[REVERSE_QUALITY_COLUMN_NAME], kmer_size=config['alignpairedend']['kmersize'])
ba = alignmentIterator(entries, aligner)
......
python/obitools3/dms/capi/kmer_similarity.pxd
View file @ e026e9ec
......@@ -32,6 +32,8 @@ cdef extern from "kmer_similarity.h" nogil:
OBIDMS_column_p column2,
index_t idx2,
index_t elt_idx2,
OBIDMS_column_p qual_col1,
OBIDMS_column_p qual_col2,
uint8_t kmer_size,
int32_t* kmer_pos_array,
int32_t* kmer_pos_array_height_p,
......
python/obitools3/dms/obiseq.pxd
View file @ e026e9ec
......@@ -5,6 +5,7 @@ from .view.view cimport Line
cdef class Seq(dict) :
cdef int _index
cpdef object clone(self)
cpdef str get_str(self)
cpdef get_symbol_at(self, int pos)
......@@ -22,6 +23,7 @@ cdef class Nuc_Seq(Seq) :
cdef class Seq_Stored(Line) :
cpdef get_symbol_at(self, int pos)
cpdef get_slice(self, slice slice_to_get)
......@@ -31,6 +33,7 @@ cdef class Nuc_Seq_Stored(Seq_Stored) :
cdef Nuc_Seq _reverse_complement
cdef object _quality_array
cdef bytes _seq
cpdef set(self, object id, object seq, object definition=*, object quality=*, int offset=*, object tags=*)
cpdef set_quality_int(self, list new_qual)
cpdef set_quality_char(self, object new_qual, int offset=*)
......
python/obitools3/dms/obiseq.pyx
View file @ e026e9ec
......@@ -40,6 +40,7 @@ cdef class Seq(dict) :
self.id = id
self.seq = seq
self.definition = definition
self._index = -1
if tags is not None :
for k in tags:
k_b = tobytes(k)
......@@ -54,6 +55,7 @@ cdef class Seq(dict) :
def new_from_stored(Seq_Stored seq_to_clone) :
cdef Seq new_seq
new_seq = Seq(seq_to_clone.id, seq_to_clone.seq, definition=seq_to_clone.definition, quality=seq_to_clone.quality, tags=seq_to_clone)
new_seq._index = seq_to_clone._index
return new_seq
def __contains__(self, object key):
......@@ -128,7 +130,16 @@ cdef class Seq(dict) :
if new_definition is not None:
new_definition = tobytes(new_definition)
self[DEFINITION_COLUMN] = new_definition
# sequence index (for reference in a view eventually) property getter and setter
@property
def index(self): # @ReservedAssignment
return self._index
@index.setter
def index(self, int idx): # @DuplicatedSignature
self._index = idx
cdef class Nuc_Seq(Seq) :
......@@ -160,6 +171,7 @@ cdef class Nuc_Seq(Seq) :
def new_from_stored(Nuc_Seq_Stored seq_to_clone) :
cdef Nuc_Seq new_seq
new_seq = Nuc_Seq(seq_to_clone.id, seq_to_clone.seq, definition=seq_to_clone.definition, quality=seq_to_clone.quality, tags=seq_to_clone)
new_seq._index = seq_to_clone.index
return new_seq
# is_revcomp property getter and setter (boolean indicating whether the sequence was created by reverse complementing another sequence)
......
python/obitools3/libalign/_solexapairend.pyx
View file @ e026e9ec
......@@ -13,7 +13,7 @@ from obitools3.dms.capi.obidmscolumn cimport OBIDMS_column_p
from obitools3.dms.view.view cimport View
from obitools3.dms.column.column cimport Column
from obitools3.commands.ngsfilter import REVERSE_SEQ_COLUMN_NAME, REVERSE_QUALITY_COLUMN_NAME
from math import log10
......@@ -182,7 +182,7 @@ def buildConsensus(ali, seq, ref_tags=None):
col_p = column.pointer()
# doesn't work because uint8_t* are forced into bytes by cython (nothing read/kept beyond 0 values)
#obi_set_qual_int_with_elt_idx_and_col_p_in_view(view_p, col_p, seq.index, 0, ali.consensus_qual, ali.consensus_len)
seq.set(ref_tags.id+b"_CONS", ali.consensus_seq, quality=ali.consensus_qual, tags=ref_tags)
seq.set(ref_tags.id+b"_CONS", ali.consensus_seq, quality=ali.consensus_qual)
seq[b'ali_length'] = ali.consensus_len
seq[b'score_norm']=float(ali.score)/ali.consensus_len
seq[b'shift']=ali.shift
......@@ -208,7 +208,7 @@ def buildConsensus(ali, seq, ref_tags=None):
quality[i] = min(42, round(-10*log10(p)))
i+=1
seq.set(ali[0].wrapped.id+b"_CONS", sseq, quality=quality, tags=ali[0].wrapped)
seq.set(ali[0].wrapped.id+b"_CONS", sseq, quality=quality)
if hasattr(ali, "counter"):
seq[b'alignement_id']=ali.counter
......@@ -224,12 +224,18 @@ def buildConsensus(ali, seq, ref_tags=None):
seq[b'ali_length']=len(seq)-ic.seqASingle-ic.seqBSingle
if seq[b'ali_length']>0:
seq[b'score_norm']=float(ali.score)/seq[b'ali_length']
ref_tags = ali[0].wrapped
if hasattr(ali, "direction"):
seq[b'ali_direction']=ali.direction
seq[b'score']=ali.score
seq[b'mode']=b'alignment'
for tag in ref_tags:
if tag != REVERSE_SEQ_COLUMN_NAME and tag != REVERSE_QUALITY_COLUMN_NAME:
seq[tag] = ref_tags[tag]
return seq
......@@ -241,10 +247,13 @@ def buildJoinedSequence(ali, reverse, seq, forward=None):
s = forward.seq + reverse.seq
quality = forward.quality
quality.extend(reverse.quality)
seq.set(forward.id +b"_PairedEnd", s, definition=forward.definition, quality=quality, tags=forward)
seq.set(forward.id +b"_PairedEnd", s, definition=forward.definition, quality=quality)
seq[b"score"]=ali.score
seq[b"ali_direction"]=ali.direction
seq[b"mode"]=b"joined"
seq[b"pairedend_limit"]=len(forward)
for tag in forward:
if tag != REVERSE_SEQ_COLUMN_NAME and tag != REVERSE_QUALITY_COLUMN_NAME:
seq[tag] = forward[tag]
return seq
python/obitools3/libalign/shifted_ali.pxd
View file @ e026e9ec
......@@ -24,7 +24,9 @@ cdef class Kmer_similarity:
cdef int32_t shift_count_array_height_a[1]
cdef Obiview_p view1_p
cdef OBIDMS_column_p column1_p
cdef OBIDMS_column_p qual_col1_p
cdef Obiview_p view2_p
cdef OBIDMS_column_p column2_p
cdef OBIDMS_column_p qual_col2_p
cdef bint build_consensus
cpdef free(self)
\ No newline at end of file
python/obitools3/libalign/shifted_ali.pyx
View file @ e026e9ec
#cython: language_level=3
from obitools3.dms.capi.obiview cimport NUC_SEQUENCE_COLUMN
from obitools3.dms.capi.obiview cimport NUC_SEQUENCE_COLUMN, QUALITY_COLUMN
from obitools3.dms.obiseq cimport Nuc_Seq_Stored
from obitools3.dms.capi.kmer_similarity cimport kmer_similarity, Obi_ali_p, obi_free_shifted_ali
from obitools3.dms.view.typed_view.view_NUC_SEQS cimport View_NUC_SEQS
from obitools3.dms.column.column cimport Column
......@@ -52,13 +51,14 @@ cdef class Ali_shifted:
def direction(self):
return self.pointer().direction
cpdef free(self):
cpdef free(self): # TODO allocated memory could be kept
obi_free_shifted_ali(self.pointer())
cdef class Kmer_similarity:
def __init__(self, View_NUC_SEQS view1, Column column1=None, View_NUC_SEQS view2=None, Column column2=None, uint8_t kmer_size=3, build_consensus=True) :
def __init__(self, View_NUC_SEQS view1, Column column1=None, Column qual_column1=None, View_NUC_SEQS view2=None, Column column2=None, Column qual_column2=None, uint8_t kmer_size=3, build_consensus=True) :
cdef Column default_seq_col
cdef Column default_qual_col
if kmer_size < 1 or kmer_size > 3:
raise Exception("Kmer size to compute kmer similarity must be >=1 or <=4")
self.kmer_pos_array_p = NULL
......@@ -89,13 +89,28 @@ cdef class Kmer_similarity:
raise Exception("Kmer similarity with no sequence column given must be given a NUC_SEQS view")
default_seq_col = view2[NUC_SEQUENCE_COLUMN]
self.column2_p = default_seq_col.pointer()
if qual_column1 is not None:
self.qual_col1_p = qual_column1.pointer()
else:
if type(view1) != View_NUC_SEQS:
raise Exception("Kmer similarity with no quality column given must be given a NUC_SEQS view")
default_qual_col = view1[QUALITY_COLUMN]
self.qual_col1_p = default_qual_col.pointer()
if qual_column2 is not None:
self.qual_col2_p = qual_column2.pointer()
else:
if type(view2) != View_NUC_SEQS:
raise Exception("Kmer similarity with no quality column given must be given a NUC_SEQS view")
default_qual_col = view2[QUALITY_COLUMN]
self.qual_col2_p = default_qual_col.pointer()
def __call__(self, Nuc_Seq_Stored seq1, Nuc_Seq_Stored seq2):
def __call__(self, object seq1, object seq2):
cdef Obi_ali_p ali_p
cdef Ali_shifted ali
ali_p = kmer_similarity(self.view1_p, self.column1_p, seq1.index, 0, \
self.view2_p, self.column2_p, seq2.index, 0, \
self.qual_col1_p, self.qual_col2_p, \
self.kmer_size, \
self.kmer_pos_array_p, self.kmer_pos_array_height_a, \
self.shift_array_p, self.shift_array_height_a, \
......
src/kmer_similarity.c
View file @ e026e9ec
......@@ -64,19 +64,18 @@ void obi_free_shifted_ali(Obi_ali_p ali)
Obi_ali_p kmer_similarity(Obiview_p view1, OBIDMS_column_p column1, index_t idx1, index_t elt_idx1,
Obiview_p view2, OBIDMS_column_p column2, index_t idx2, index_t elt_idx2,
uint8_t kmer_size,
int32_t* kmer_pos_array, int32_t* kmer_pos_array_height_p,
int32_t* shift_array, int32_t* shift_array_height_p,
int32_t* shift_count_array, int32_t* shift_count_array_length_p,
bool build_consensus)
Obiview_p view2, OBIDMS_column_p column2, index_t idx2, index_t elt_idx2,
OBIDMS_column_p qual_col1, OBIDMS_column_p qual_col2,
uint8_t kmer_size,
int32_t* kmer_pos_array, int32_t* kmer_pos_array_height_p,
int32_t* shift_array, int32_t* shift_array_height_p,
int32_t* shift_count_array, int32_t* shift_count_array_length_p,
bool build_consensus)
{
Obi_ali_p ali = NULL;
int i, j;
bool switched_seqs;
bool left_ali;
OBIDMS_column_p qual_col1;
OBIDMS_column_p qual_col2;
double score = 0.0;
Obi_blob_p blob1 = NULL;
Obi_blob_p blob2 = NULL;
......@@ -106,6 +105,8 @@ Obi_ali_p kmer_similarity(Obiview_p view1, OBIDMS_column_p column1, index_t idx1
int32_t overlap_len;
int32_t consensus_len;
int32_t cons_shift;
int32_t copy_start;
int32_t copy_len;
uint8_t kmer;
byte_t nuc;
uint8_t encoding;
......@@ -116,6 +117,10 @@ Obi_ali_p kmer_similarity(Obiview_p view1, OBIDMS_column_p column1, index_t idx1
int32_t shift_array_height = *shift_array_height_p;
int32_t shift_count_array_length = *shift_count_array_length_p;
bool free_blob1 = false;
bool keep_seq1_start;
bool keep_seq2_start;
bool keep_seq1_end;
bool keep_seq2_end;
// Check kmer size
if ((kmer_size < 1) || (kmer_size > 4))
......@@ -143,7 +148,7 @@ Obi_ali_p kmer_similarity(Obiview_p view1, OBIDMS_column_p column1, index_t idx1
switched_seqs = false;
len1 = blob1->length_decoded_value;
len2 = blob2->length_decoded_value;
if (len2 > len1)
if (len2 < len1)
{
switched_seqs = true;
temp_len = len1;
......@@ -334,7 +339,8 @@ Obi_ali_p kmer_similarity(Obiview_p view1, OBIDMS_column_p column1, index_t idx1
// Find the most represented shift
best_shift_idx = 0;
max_common_kmers = 0;
empty_part = (shift_count_array_length-1)/2 - len1;
//empty_part = (shift_count_array_length-1)/2 - len1; //TODO wrong in some cases (len1 shorter than overlap or something like that)
empty_part = 0;
for (i=empty_part; i < (shift_count_array_length - empty_part); i++) // skipping empty parts of the array
{
if (shift_count_array[i] > max_common_kmers)
......@@ -345,10 +351,44 @@ Obi_ali_p kmer_similarity(Obiview_p view1, OBIDMS_column_p column1, index_t idx1
}
best_shift = best_shift_idx - len1;
keep_seq1_start = false;
keep_seq1_end = false;
keep_seq2_start = false;
keep_seq2_end = false;
// The 873863 cases of hell
if (best_shift > 0)
overlap_len = len2 - best_shift;
else
{
overlap_len = len2 - best_shift;
if (len1 <= overlap_len)
{
overlap_len = len1;
if (! switched_seqs)
keep_seq2_end = true;
else
keep_seq2_start = true;
}
else if (switched_seqs)
{
keep_seq2_start = true;
keep_seq1_end = true;
}
}
else if (best_shift < 0)
{
overlap_len = len1 + best_shift;
if (!switched_seqs)
{
keep_seq1_start = true;
keep_seq2_end = true;
}
}
else
{
overlap_len = len1;
if ((!switched_seqs) && (len2 > len1))
keep_seq2_end = true;
}
score = max_common_kmers + kmer_size - 1;
......@@ -369,7 +409,7 @@ Obi_ali_p kmer_similarity(Obiview_p view1, OBIDMS_column_p column1, index_t idx1
ali->shift = abs_shift;
ali->consensus_seq = NULL;
ali->consensus_qual = NULL;
if (((best_shift < 0) && (!switched_seqs)) || ((best_shift > 0) && switched_seqs))
if (((best_shift <= 0) && (!switched_seqs)) || ((best_shift > 0) && switched_seqs))
{
left_ali = true;
strcpy(ali->direction, "left");
......@@ -383,33 +423,17 @@ Obi_ali_p kmer_similarity(Obiview_p view1, OBIDMS_column_p column1, index_t idx1
// Build the consensus sequence if asked
if (build_consensus)
{
// Get the quality columns
qual_col1 = obi_view_get_column(view1, QUALITY_COLUMN);
if (qual_col1 == NULL)
{
obi_set_errno(OBI_ALIGN_ERROR);
obidebug(1, "\nError when computing the kmer similarity between two sequences: can't open quality column to build the consensus sequence");
return NULL;
}
qual_col2 = obi_view_get_column(view2, QUALITY_COLUMN);
if (qual_col2 == NULL)
{
obi_set_errno(OBI_ALIGN_ERROR);
obidebug(1, "\nError when computing the kmer similarity between two sequences: can't open quality column to build the consensus sequence");
return NULL;
}
// Get the quality arrays
qual1 = obi_get_qual_int_with_elt_idx_and_col_p_in_view(view1, qual_col1, idx1, 0, &qual1_len);
if (qual1 == NULL)
{
obidebug(1, "\nError getting the blob of the 1st sequence when computing the kmer similarity between two sequences");
obidebug(1, "\nError getting the quality of the 1st sequence when computing the kmer similarity between two sequences");
return NULL;
}
qual2 = obi_get_qual_int_with_elt_idx_and_col_p_in_view(view2, qual_col2, idx2, 0, &qual2_len);
if (qual2 == NULL)
{
obidebug(1, "\nError getting the blob of the 1st sequence when computing the kmer similarity between two sequences");
obidebug(1, "\nError getting the quality of the 2nd sequence when computing the kmer similarity between two sequences");
return NULL;
}
......@@ -417,10 +441,10 @@ Obi_ali_p kmer_similarity(Obiview_p view1, OBIDMS_column_p column1, index_t idx1
if (seq1 == NULL)
seq1 = obi_blob_to_seq(blob1);
if (left_ali)
consensus_len = len1 + len2 - overlap_len;
if (! switched_seqs)
consensus_len = len2 - best_shift;
else
consensus_len = overlap_len;
consensus_len = len1 + best_shift;
// Allocate memory for consensus sequence
consensus_seq = (char*) malloc(consensus_len + 1 * sizeof(char)); // TODO keep malloced too maybe
......@@ -447,30 +471,27 @@ Obi_ali_p kmer_similarity(Obiview_p view1, OBIDMS_column_p column1, index_t idx1
// Compute consensus-relative shift for each sequence
if (best_shift > 0)
{
shift1 = best_shift;
shift2 = 0;
shift1 = 0;
shift2 = best_shift;
}
else
{
shift1 = 0;
shift2 = -(best_shift);
shift1 = -(best_shift);
shift2 = 0;
}
// If left alignment, copy first part of first sequence
if (left_ali)
// Copy first part of first or second sequence depending on cases
if (keep_seq1_start)
{
if (switched_seqs)
{
strncpy(consensus_seq, seq2, abs_shift);
memcpy(consensus_qual, qual2, abs_shift*sizeof(uint8_t));
cons_shift = abs_shift;
}
else
{
strncpy(consensus_seq, seq1, abs_shift);
memcpy(consensus_qual, qual1, abs_shift*sizeof(uint8_t));
cons_shift = abs_shift;
}
strncpy(consensus_seq, seq1, abs_shift);
memcpy(consensus_qual, qual1, abs_shift*sizeof(uint8_t));
cons_shift = abs_shift;
}
else if (keep_seq2_start)
{
strncpy(consensus_seq, seq2, abs_shift);
memcpy(consensus_qual, qual2, abs_shift*sizeof(uint8_t));
cons_shift = abs_shift;
}
else
cons_shift = 0;
......@@ -485,19 +506,26 @@ Obi_ali_p kmer_similarity(Obiview_p view1, OBIDMS_column_p column1, index_t idx1
consensus_qual[pos+cons_shift] = round((qual1[pos+shift1] + qual2[pos+shift2])/2); // TODO maybe use the (p1*(1-p2/3)) formula (but parenthesis bug???)
}
// If left alignment, copy last part of first sequence
if (left_ali)
// Copy last part of first or second sequence depending on cases
if (keep_seq1_end)
{
strncpy(consensus_seq+cons_shift+overlap_len, seq1+overlap_len, len1 - overlap_len);
memcpy(consensus_qual+cons_shift+overlap_len, qual1+overlap_len, (len1 - overlap_len)*sizeof(uint8_t));
}
if (keep_seq2_end)
{
if (switched_seqs)
if (best_shift <= 0)
{
strncpy(consensus_seq+cons_shift+overlap_len, seq1+overlap_len, len1-overlap_len);
memcpy(consensus_qual+cons_shift+overlap_len, qual1+overlap_len, (len1-overlap_len)*sizeof(uint8_t));
copy_start = overlap_len;
copy_len = len2 - overlap_len;
}
else
if (best_shift > 0)
{
strncpy(consensus_seq+cons_shift+overlap_len, seq2+overlap_len, len2-overlap_len);
memcpy(consensus_qual+cons_shift+overlap_len, qual2+overlap_len, (len2-overlap_len)*sizeof(uint8_t));
copy_start = overlap_len + best_shift;
copy_len = len2 - overlap_len - best_shift;
}
strncpy(consensus_seq+cons_shift+overlap_len, seq2+copy_start, copy_len);
memcpy(consensus_qual+cons_shift+overlap_len, qual2+copy_start, copy_len*sizeof(uint8_t));
}
consensus_seq[consensus_len] = '\0';
......
src/kmer_similarity.h
View file @ e026e9ec
......@@ -33,9 +33,9 @@ typedef struct Obi_ali {
*/
int overlap_length; /**< Length of the overlap between the aligned sequences.
*/
char* consensus_seq; /**< Consensus sequence built with the computed overlap.
char* consensus_seq; /**< Consensus sequence built as to reconstruct a pairedend read.
*/
uint8_t* consensus_qual; /**< Consensus quality built with the computed overlap.
uint8_t* consensus_qual; /**< Consensus quality built as to reconstruct a pairedend read.
*/
int shift; /**< Shift chosen to align the sequences (for shifted alignment).
*/
......@@ -91,7 +91,7 @@ void obi_free_shifted_ali(Obi_ali_p ali);
* If needed, reallocated to a bigger size.
* @param shift_count_array_height_p A pointer on an integer corresponding to the size (number of elements)
* allocated for shift_count_array. Updated by the function as needed.
* @param build_consensus A boolean indicating whether the function should build the consensus sequence and quality. // TODO option to build consensus without quality?
* @param build_consensus A boolean indicating whether the function should build the consensus sequence and quality as to reconstruct a pairedend read. // TODO option to build consensus without quality?
*
* @returns A pointer on an Obi_ali_p structure containing the results.
* @retval NULL if an error occurred.
......@@ -107,6 +107,8 @@ Obi_ali_p kmer_similarity(Obiview_p view1,
OBIDMS_column_p column2,
index_t idx2,
index_t elt_idx2,
OBIDMS_column_p qual_col1,
OBIDMS_column_p qual_col2,
uint8_t kmer_size,
int32_t* kmer_pos_array,
int32_t* kmer_pos_array_height_p,
......
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