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/*
* filters.cpp
* cufflinks
*
* Created by Cole Trapnell on 10/27/09.
* Copyright 2009 Cole Trapnell. All rights reserved.
*
*/
#include "filters.h"
#include <algorithm>
#include <numeric>
#include <boost/graph/adjacency_list.hpp>
#include <boost/graph/depth_first_search.hpp>
#include <boost/graph/visitors.hpp>
#include <boost/graph/graph_traits.hpp>
#include <boost/graph/connected_components.hpp>
using namespace std;
using namespace boost;
void filter_introns(int bundle_length,
int bundle_left,
vector<Scaffold>& hits,
double fraction,
bool filter_on_intron_overlap,
bool filter_with_intron_doc)
{
vector<float> depth_of_coverage(bundle_length,0);
vector<double> scaff_doc;
map<pair<int,int>, float> intron_doc;
vector<Scaffold> filtered_hits;
vector<bool> toss(hits.size(), false);
double bundle_avg_doc = compute_doc(bundle_left,
hits,
depth_of_coverage,
intron_doc,
false);
double bundle_avg_thresh = bundle_avg_doc * fraction;
if (filter_with_intron_doc && !intron_doc.empty())
{
bundle_avg_doc = major_isoform_intron_doc(intron_doc);
bundle_avg_thresh = fraction * bundle_avg_doc;
verbose_msg("\tFiltering bundle introns, avg (intron) doc = %lf, thresh = %f\n", bundle_avg_doc, bundle_avg_thresh);
}
else
{
verbose_msg("\tFiltering bundle introns, avg bundle doc = %lf, thresh = %f\n", bundle_avg_doc, bundle_avg_thresh);
}
for(map<pair<int, int>, float>::const_iterator itr = intron_doc.begin();
itr != intron_doc.end();
++itr)
{
for (size_t j = 0; j < hits.size(); ++j)
{
//fprintf(stderr, "considering read [%d-%d] with min doc = %lf contained in intron with doc = %lf\n", hits[j].left(), hits[j].right(), doc, idoc);
const vector<AugmentedCuffOp>& ops = hits[j].augmented_ops();
for (size_t i = 0; i < ops.size(); ++i)
{
if (ops[i].opcode == CUFF_INTRON)
{
map<pair<int, int>, float>::const_iterator itr;
itr = intron_doc.find(make_pair(ops[i].g_left(), ops[i].g_right()));
double doc = itr->second;
if (doc < bundle_avg_thresh)
{
toss[j] = true;
verbose_msg("\t Filtering intron %d - %d: %f thresh %f\n", itr->first.first, itr->first.second, doc, bundle_avg_thresh);
continue;
}
if (!filter_on_intron_overlap)
continue;
for (map<pair<int,int>, float>::const_iterator itr2 = intron_doc.begin();
itr2 != intron_doc.end();
++itr2)
{
if (itr == itr2 ||
!overlap_in_genome(itr->first.first,
itr->first.second,
itr2->first.first,
itr2->first.second))
continue;
double thresh = itr2->second * fraction;
if (doc < thresh)
{
verbose_msg("\t Filtering intron (due to overlap) %d - %d: %f thresh %f\n", itr->first.first, itr->first.second, doc, bundle_avg_thresh);
toss[j] = true;
}
}
}
}
}
}
for (size_t j = 0; j < hits.size(); ++j)
{
if (!toss[j])
{
filtered_hits.push_back(hits[j]);
//#if verbose_msg
// if (hits[j].has_intron())
// {
// fprintf(stderr, "KEEPING intron scaff [%d-%d]\n", hits[j].left(), hits[j].right());
// }
//#endif
}
else
{
if (hits[j].has_intron())
{
verbose_msg("\tFiltering intron scaff [%d-%d]\n", hits[j].left(), hits[j].right());
}
}
}
verbose_msg("\tIntron filtering pass finished: excluded %d fragments\n", (int)hits.size() - (int)filtered_hits.size());
hits = filtered_hits;
}
double background_rate(const vector<float> depth_of_coverage,
int left,
int right)
{
vector<float> tmp;
size_t r_bound = (size_t)min(right, (int) depth_of_coverage.size());
size_t l_bound = (size_t)max(left, 0);
tmp.insert(tmp.end(),
depth_of_coverage.begin() + l_bound,
depth_of_coverage.begin() + r_bound);
if (tmp.empty())
return 0;
vector<float>::iterator new_end = remove(tmp.begin(), tmp.end(), 0);
tmp.erase(new_end, tmp.end());
sort(tmp.begin(), tmp.end());
size_t median = (size_t)floor(tmp.size() / 2);
double median_doc = tmp[median];
return median_doc;
}
void pre_mrna_filter(int bundle_length,
int bundle_left,
vector<Scaffold>& hits)
{
vector<float> depth_of_coverage(bundle_length,0);
vector<double> scaff_doc;
map<pair<int,int>, float> intron_doc;
vector<Scaffold> filtered_hits;
vector<bool> toss(hits.size(), false);
vector<float> through_introns; //for each location, how many introns pass through
vector<int> scaff_intron_status;
// Make sure the avg only uses stuff we're sure isn't pre-mrna fragments
double bundle_avg_doc = compute_doc(bundle_left,
hits,
depth_of_coverage,
intron_doc,
true,
&through_introns,
&scaff_intron_status);
verbose_msg("Pre-mRNA flt: bundle average doc = %lf\n", bundle_avg_doc);
/*
//2nd call not needed, the vectors won't change, only the return value
compute_doc(bundle_left,
hits,
depth_of_coverage,
intron_doc,
false);
*/
record_doc_for_scaffolds(bundle_left,
hits,
depth_of_coverage,
intron_doc,
scaff_doc);
for(map<pair<int, int>, float >::const_iterator itr = intron_doc.begin();
itr != intron_doc.end();
++itr)
{
int i_left = itr->first.first;
int i_right = itr->first.second;
int i_doc = itr->second;
double intron_background = background_rate(depth_of_coverage,
i_left - bundle_left,
i_right - bundle_left);
double cumul_cov = 0;
for (int i = 0; i < i_right - i_left; ++i)
{
size_t pos = (i_left - bundle_left) + i;
cumul_cov += depth_of_coverage[pos];
}
cumul_cov /= i_right - i_left;
verbose_msg("Pre-mRNA flt: intron %d-%d : background: %lf, inner coverage: %lf, junction coverage: %f\n",
i_left, i_right, intron_background, cumul_cov, i_doc);
if (cumul_cov / bundle_avg_doc >= pre_mrna_fraction)
{
//fprintf(stderr, "\tskipping\n");
continue;
}
////double thresh = (1.0/pre_mrna_fraction) * intron_background;
double thresh = pre_mrna_fraction * intron_background;
float min_flt_fraction = min(pre_mrna_fraction, min_isoform_fraction);
//double thresh = min_flt_fraction * i_doc;
for (size_t j = 0; j < hits.size(); ++j)
{
if (hits[j].left()>i_right) break;
if (hits[j].is_ref())
continue;
if (toss[j])
continue;
//find maximum intron support in the hit region
int len = 0;
double doc = 0.0;
size_t curr_op = 0;
const vector<AugmentedCuffOp>& ops = hits[j].augmented_ops();
while (curr_op != ops.size())
{
const AugmentedCuffOp& op = ops[curr_op];
if (op.opcode == CUFF_MATCH)
{
int op_len = 0;
double op_doc = 0.0;
int left_off = op.g_left();
if (left_off + op.genomic_length > i_left && left_off < i_right)
{
if (left_off > i_left)
{
if (left_off + op.genomic_length <= i_right + 1)
{
op_len += op.genomic_length;
int L = left_off - bundle_left;
int R = L + op.genomic_length;
op_doc += accumulate(depth_of_coverage.begin() + L, depth_of_coverage.begin() + R, 0);
}
else
{
op_len += i_right - left_off;
int L = left_off - bundle_left;
int R = L + (i_right - left_off);
op_doc += accumulate(depth_of_coverage.begin() + L, depth_of_coverage.begin() + R, 0);
}
}
else
{
if (left_off + op.genomic_length <= i_right + 1)
{
op_len += (left_off + op.genomic_length - i_left);
int L = left_off - bundle_left;
int R = L + (left_off + op.genomic_length - i_left);
op_doc += accumulate(depth_of_coverage.begin() + L, depth_of_coverage.begin() + R, 0);
}
else
{
op_len = i_right - i_left;
int L = left_off - bundle_left;
int R = L + (i_right - i_left);
op_doc = accumulate(depth_of_coverage.begin() + L, depth_of_coverage.begin() + R, 0);
}
}
}
len += op_len;
doc += op_doc;
}
if (op.g_left() >= i_right)
break;
++curr_op;
}
if (len)
{
double hit_doc_in_region = doc / len;
if (hit_doc_in_region < thresh)
{
toss[j] = true;
if (hits[j].has_intron())
{
// fprintf(stderr, "\t$$$ Filtering intron scaff [%d-%d]\n", hits[j].left(), hits[j].right());
verbose_msg("\t@@@ Filtering intron scaff [%d-%d] (scaff_doc=%lf, doc_in_region=%lf)\n",
hits[j].left(), hits[j].right(), scaff_doc[j], hit_doc_in_region);
}
}
}
} //for each scaffold
} //for each intron
for (size_t j = 0; j < hits.size(); ++j)
{
if (!toss[j])
{
filtered_hits.push_back(hits[j]);
}
/*else
{
if (hits[j].has_intron())
{
verbose_msg( "\t@@@ Filtering intron scaff [%d-%d]\n", hits[j].left(), hits[j].right());
}
}
*/
}
if (cuff_verbose && hits.size()>filtered_hits.size())
verbose_msg("\tPre-mRNA flt tossed %lu fragments\n", hits.size() - filtered_hits.size());
hits = filtered_hits;
}
void filter_hits(int bundle_length,
int bundle_left,
vector<Scaffold>& hits)
{
pre_mrna_filter(bundle_length, bundle_left, hits);
vector<float> depth_of_coverage(bundle_length+1,0);
vector<double> scaff_doc;
map<pair<int,int>, float> intron_doc;
vector<Scaffold> filtered_hits;
vector<bool> toss(hits.size(), false);
// Make sure the avg only uses stuff we're sure isn't pre-mrna fragments
double bundle_avg_doc = compute_doc(bundle_left,
hits,
depth_of_coverage,
intron_doc,
true);
// recompute the real DoCs
/* not needed, vectors are not changed
compute_doc(bundle_left,
hits,
depth_of_coverage,
intron_doc,
false);
*/
record_min_doc_for_scaffolds(bundle_left,
hits,
depth_of_coverage,
intron_doc,
scaff_doc);
//double bundle_avg_thresh = min_isoform_fraction * bundle_avg_doc;
if (!intron_doc.empty())
{
double intron_avg_doc = major_isoform_intron_doc(intron_doc);
double intron_multiplier = intron_avg_doc / bundle_avg_doc;
// we don't want this to be more than 1.0 ...
intron_multiplier = min(intron_avg_doc, 1.0);
//bundle_avg_thresh = min_isoform_fraction * bundle_avg_doc;
set<pair<int, int> > tossed_introns;
for(map<pair<int, int>, float>::const_iterator itr = intron_doc.begin();
itr != intron_doc.end();
++itr)
{
for (size_t j = 0; j < hits.size(); ++j)
{
if (hits[j].is_ref())
{
continue;
}
int i_left = itr->first.first;
int i_right = itr->first.second;
int j_match_len = hits[j].match_length(i_left, i_right);
if (j_match_len > 0)
{
double idoc = itr->second;
double doc = scaff_doc[j];
if (!hits[j].has_intron() &&
doc < pre_mrna_fraction * (idoc * intron_multiplier))
{
toss[j] = true;
}
const vector<AugmentedCuffOp>& ops = hits[j].augmented_ops();
unsigned int num_mismatches = 0;
assert (hits[j].mate_hits().size() == 1);
const MateHit& hit = **(hits[j].mate_hits().begin());
num_mismatches = hit.edit_dist();
double percent_mismatches = num_mismatches / (double)hits[j].length();
bool intron_pokin_read = false;
const AugmentedCuffOp& first = ops.front();
// intron =========================
// hit ******************
if (first.g_left() < i_left && first.g_right() > i_left && first.g_right() < i_right)
{
intron_pokin_read = true;
}
// intron =========================
// hit ******************
if (first.g_left() < i_right && first.g_right() > i_right && first.g_left() > i_left)
{
intron_pokin_read = true;
}
const AugmentedCuffOp& last = ops.back();
// intron =========================
// hit ******************
if (last.g_left() < i_left && last.g_right() > i_left && last.g_right() < i_right)
{
intron_pokin_read = true;
}
// intron =========================
// hit ******************
if (last.g_left() < i_right && last.g_right() > i_right && last.g_left() > i_left)
{
intron_pokin_read = true;
}
if (intron_pokin_read)
{
double fraction;
// if (!hits[j].has_intron())
// {
// fraction = (3 * pre_mrna_fraction) + percent_mismatches;
// }
// else
{
fraction = pre_mrna_fraction + percent_mismatches;
}
double thresh = fraction * (intron_avg_doc * intron_multiplier);
if (doc < thresh)
{
toss[j] = true;
// if (hits[j].has_intron())
// {
// fprintf(stderr, "\t^^^Filtering intron scaff [%d-%d]\n", hits[j].left(), hits[j].right());
// }
}
}
}
}
}
}
for (size_t j = 0; j < hits.size(); ++j)
{
if (!toss[j])
{
filtered_hits.push_back(hits[j]);
//#if verbose_msg
// if (hits[j].has_intron())
// {
//
// fprintf(stderr, "KEEPING intron scaff [%d-%d]\n", hits[j].left(), hits[j].right());
// }
//#endif
}
else
{
if (hits[j].has_intron())
{
verbose_msg("\t!!!Filtering intron scaff [%d-%d]\n", hits[j].left(), hits[j].right());
}
}
}
//#if verbose_msg
// fprintf(stderr, "\tInitial filter pass complete\n");
//#endif
hits = filtered_hits;
scaff_doc.clear();
filtered_hits.clear();
toss = vector<bool>(hits.size(), false);
map<pair<int, int>, float> dummy;
bundle_avg_doc = compute_doc(bundle_left,
hits,
depth_of_coverage,
dummy,
false);
//#if verbose_msg
// fprintf(stderr, "\tUpdated avg bundle doc = %lf\n", bundle_avg_doc);
//#endif
record_doc_for_scaffolds(bundle_left,
hits,
depth_of_coverage,
intron_doc,
scaff_doc);
//#if verbose_msg
// double bundle_thresh = pre_mrna_fraction * bundle_avg_doc;
// fprintf(stderr, "\tthreshold is = %lf\n", bundle_thresh);
//#endif
if (!intron_doc.empty())
{
// filter_introns(bundle_length,
// bundle_left,
// hits,
// min_isoform_fraction,
// true,
// true);
if (bundle_avg_doc > 3000)
{
filter_introns(bundle_length,
bundle_left,
hits,
min_isoform_fraction,
true,
false);
}
}
for (size_t j = 0; j < hits.size(); ++j)
{
if (!toss[j])
{
filtered_hits.push_back(hits[j]);
//#if verbose_msg
// if (hits[j].has_intron())
// {
//
// fprintf(stderr, "KEEPING intron scaff [%d-%d]\n", hits[j].left(), hits[j].right());
// }
//#endif
}
else
{
if (hits[j].has_intron())
{
verbose_msg("\t***Filtering intron scaff [%d-%d]\n", hits[j].left(), hits[j].right());
}
}
}
//fprintf(stderr, "\tTossed %d hits as noise\n", (int)hits.size() - filtered_hits.size());
hits = filtered_hits;
}
void filter_junk_isoforms(vector<shared_ptr<Abundance> >& transcripts,
vector<double>& abundances,
const vector<shared_ptr<Abundance> >& mapped_transcripts,
double locus_mass)
{
// vector<double>::iterator max_ab = std::max_element(abundances.begin(),
// abundances.end());
double max_fwd_ab = -1.0;
double max_rev_ab = -1.0;
for (size_t t = 0; t < transcripts.size(); ++t)
{
shared_ptr<Scaffold> scaff = transcripts[t]->transfrag();
if (scaff->strand() == CUFF_FWD || scaff->strand() == CUFF_STRAND_UNKNOWN)
{
if (abundances[t] > max_fwd_ab)
max_fwd_ab = abundances[t];
}
if (scaff->strand() == CUFF_REV || scaff->strand() == CUFF_STRAND_UNKNOWN)
{
if (abundances[t] > max_rev_ab)
max_rev_ab = abundances[t];
}
}
// Try to categorize the crap transcripts for suppression
vector<bool> pre_mrna_junk(transcripts.size(), false); //intra-intron, much lower abundance than container
vector<bool> chaff(transcripts.size(), false); // only a single MateHit, impossible to reliably quantitate
vector<bool> repeats(transcripts.size(), false); // too many low-quality hits
vector<bool> too_rare(transcripts.size(), false); // too rare to be reliably quantitated, could be error
//cerr << "Chucked : ";
for (size_t t = 0; t < transcripts.size(); ++t)
{
shared_ptr<Scaffold> scaff = transcripts[t]->transfrag();
if (!(scaff->is_ref()) && allow_junk_filtering)
{
const vector<const MateHit*> hits = scaff->mate_hits();
const vector<AugmentedCuffOp>& ops = scaff->augmented_ops();
if (ops.size() == 1 && ops[0].opcode == CUFF_MATCH)
{
for (size_t j = 0; j < transcripts.size(); ++j)
{
const vector<AugmentedCuffOp>& j_ops = scaff->augmented_ops();
for (size_t L = 0; L < j_ops.size(); L++)
{
if (AugmentedCuffOp::overlap_in_genome(ops[0], j_ops[L]) &&
j_ops[L].opcode == CUFF_INTRON)
{
pre_mrna_junk[t] = true;
}
}
}
}
if (library_type != "transfrags")
{
double low_qual_hits = 0.0;
static const double low_qual_err_prob = high_phred_err_prob; // hits with error_prob() above this are low quality;
static const double low_qual_thresh = 0.75; // hits with more than this fraction of low qual hits are repeats
for (vector<const MateHit*>::const_iterator itr = hits.begin();
itr != hits.end();
++itr)
{
double e = 1-(*itr)->mass();
if (e >= low_qual_err_prob)
low_qual_hits += 1.0;
}
double low_qual_frac = low_qual_hits / (double)hits.size();
if (low_qual_frac > low_qual_thresh)
repeats[t] = true;
}
if (scaff->strand() == CUFF_FWD &&
(abundances[t] / max_fwd_ab) < min_isoform_fraction)
too_rare[t] = true;
if ((scaff->strand() == CUFF_REV || scaff->strand() == CUFF_STRAND_UNKNOWN) &&
(abundances[t] / max_rev_ab) < min_isoform_fraction)
too_rare[t] = true;
const vector<double>* cond_probs = (mapped_transcripts[t]->cond_probs());
if (cond_probs)
{
assert (library_type != "transfrags");
double supporting_hits = abundances[t] * locus_mass;
if (supporting_hits < min_frags_per_transfrag)
chaff[t] = true;
}
}
// else // we should still filter things that are zero to improve robustness of MAP estimation
// {
// if (abundances[t] == 0.0)
// too_rare[t] = true;
// }
}
vector<shared_ptr<Abundance> > non_junk_transcripts;
vector<double> non_junk_abundances;
for (size_t t = 0; t < transcripts.size(); ++t)
{
if (!repeats[t] && !pre_mrna_junk[t] && !too_rare[t] && !chaff[t])
{
non_junk_transcripts.push_back(transcripts[t]);
non_junk_abundances.push_back(abundances[t]);
}
else
{
verbose_msg( "Filtering isoform %d-%d\n", transcripts[t]->transfrag()->left(), transcripts[t]->transfrag()->right());
}
}
transcripts = non_junk_transcripts;
abundances = non_junk_abundances;
}
// Designed to strip out remaining pre-mrna genes, assembled repeats, and
// fragments from isoforms too short to be reliably quantitated.
void filter_junk_genes(vector<Gene>& genes)
{
vector<Gene> good_genes;
vector<Isoform> all_isoforms;
for (size_t i = 0; i < genes.size(); ++i)
{
all_isoforms.insert(all_isoforms.end(),
genes[i].isoforms().begin(),
genes[i].isoforms().end());
}
for (size_t i = 0; i < genes.size(); ++i)
{
const Gene& g = genes[i];
if(g.has_ref_trans())
{
good_genes.push_back(g);
continue;
}
bool good_gene = true;
for (size_t j = 0; j < all_isoforms.size(); ++j)
{
vector<pair<int, int> > introns = all_isoforms[j].scaffold().gaps();
//assert (!allow_junk_filtering || all_isoforms[j].scaffold().mate_hits().size() >= min_frags_per_transfrag);
for (size_t k = 0; k < introns.size(); ++k)
{
if (g.left() > introns[k].first && g.right() < introns[k].second &&
g.FPKM() / all_isoforms[j].FPKM() < pre_mrna_fraction)
{
good_gene = false;
}
}
}
if (allow_junk_filtering)
{
if (g.FPKM() == 0)
{
good_gene = false;
}
}
if (good_gene)
{
good_genes.push_back(g);
}
else
{
verbose_msg("Filtering transfrags from gene %d-%d\n", g.left(), g.right());
}
}
genes = good_genes;
}
void clip_by_3_prime_dropoff(vector<Scaffold>& scaffolds)
{
vector<pair<double, Scaffold*> > three_prime_ends;
if (library_type != "transfrags")
{
foreach (Scaffold& scaff, scaffolds)
{
if (!(scaff.strand() == CUFF_FWD || scaff.strand() == CUFF_REV))
continue;
int scaff_len = scaff.length();
vector<double> coverage(scaff_len, 0.0);
double total = 0;
foreach(const MateHit* hit, scaff.mate_hits())
{
int start, end, frag_len;
if (!scaff.map_frag(*hit, start, end, frag_len)) continue;
if (scaff.strand() == CUFF_REV)
{
start = scaff_len - 1 - start;
end = scaff_len - 1 - end;
swap(start, end);
}
for(int i = start; i <= end; ++i)
{
coverage[i] += hit->mass();
total += hit->mass();
}
}
double avg_cov = total/scaff_len;
// if (avg_cov < trim_3_avgcov_thresh)
// continue;
const AugmentedCuffOp* exon_3 = NULL;
int mult;
int offset;
if (scaff.strand() == CUFF_REV)
{
mult = 1;
offset = 0;
exon_3 = &scaff.augmented_ops().front();
}
else if (scaff.strand() == CUFF_FWD)
{
mult = -1;
offset = scaff_len - 1;
exon_3 = &scaff.augmented_ops().back();
}
else
{
continue;
}
int to_remove;
double min_cost = numeric_limits<double>::max();
double mean_to_keep = 0.0;
double mean_to_trim = 0.0;
double tmp_mean_to_trim = 0.0;
double tmp_mean_to_keep = 0.0;
double tmp_mean_3prime = 0.0;
for (int i = 0; i < exon_3->genomic_length; i++)
{
tmp_mean_3prime += coverage[offset + mult*i];
}
tmp_mean_3prime /= exon_3->genomic_length;
double base_cost = 0.0;
for (int i = 0; i < exon_3->genomic_length; i++)
{
double d = (coverage[offset + mult*i] - tmp_mean_3prime);
d *= d;
base_cost += d;
}
base_cost /= exon_3->genomic_length;
size_t min_cost_x = -1;
for (to_remove = 1; to_remove < exon_3->genomic_length - 1; to_remove++)
{
tmp_mean_to_trim = 0.0;
tmp_mean_to_keep = 0.0;
for (size_t i = 0; i < exon_3->genomic_length; i++)
{
if (i <= to_remove)
{
tmp_mean_to_trim += coverage[offset + mult*i];
}
else
{
tmp_mean_to_keep += coverage[offset + mult*i];
}
}
tmp_mean_to_trim /= to_remove;
tmp_mean_to_keep /= (exon_3->genomic_length - to_remove);
double tmp_mean_trim_cost = 0.0;
double tmp_mean_keep_cost = 0.0;
for (int i = 0; i < exon_3->genomic_length; i++)
{
if (i <= to_remove)
{
double d = (coverage[offset + mult*i] - tmp_mean_to_trim);
d *= d;
tmp_mean_trim_cost += d;
}
else
{
double d = (coverage[offset + mult*i] - tmp_mean_to_keep);
d *= d;
tmp_mean_keep_cost += d;
}
}
tmp_mean_trim_cost /= to_remove;
tmp_mean_keep_cost /= (exon_3->genomic_length - to_remove);
double new_cost = tmp_mean_trim_cost + tmp_mean_keep_cost;
if (new_cost < min_cost && trim_3_dropoff_frac * tmp_mean_to_keep > tmp_mean_to_trim && new_cost < base_cost && to_remove > scaff_len * 0.05)
{
min_cost = tmp_mean_trim_cost + tmp_mean_keep_cost;
min_cost_x = to_remove;
mean_to_keep = tmp_mean_to_keep;
mean_to_trim = tmp_mean_to_trim;
}
}
// If trimming reduces the overall mean squared error of the coverage
// do it
if (avg_cov >= trim_3_avgcov_thresh && min_cost_x < exon_3->genomic_length)
{
scaff.trim_3(min_cost_x);
}
// store the mean squared error for this exon
tmp_mean_3prime = 0.0;
for (int i = 0; i < exon_3->genomic_length; i++)
{
tmp_mean_3prime += coverage[offset + mult*i];
}
tmp_mean_3prime /= exon_3->genomic_length;
base_cost = 0.0;
for (int i = 0; i < exon_3->genomic_length; i++)
{
double d = (coverage[offset + mult*i] - tmp_mean_3prime);
d *= d;
base_cost += d;
}
base_cost /= exon_3->genomic_length;
three_prime_ends.push_back(make_pair(base_cost, &scaff));
}
}
else
{
foreach (Scaffold& scaff, scaffolds)
{
if (!(scaff.strand() == CUFF_FWD || scaff.strand() == CUFF_REV))
continue;
int scaff_len = scaff.length();
vector<double> coverage(scaff_len, 0.0);
double total = 0;
foreach(const MateHit* hit, scaff.mate_hits())
{
int start, end, frag_len;
if (!scaff.map_frag(*hit, start, end, frag_len)) continue;
if (scaff.strand() == CUFF_REV)
{
start = scaff_len - 1 - start;
end = scaff_len - 1 - end;
swap(start, end);
}
for(int i = start; i <= end; ++i)
{
coverage[i] += hit->mass();
total += hit->mass();
}
}
double avg_cov = total/scaff_len;
// if (avg_cov < trim_3_avgcov_thresh)
// continue;
const AugmentedCuffOp* exon_3 = NULL;
int mult;
int offset;
if (scaff.strand() == CUFF_REV)
{
mult = 1;
offset = 0;
exon_3 = &scaff.augmented_ops().front();
}
else if (scaff.strand() == CUFF_FWD)
{
mult = -1;
offset = scaff_len - 1;
exon_3 = &scaff.augmented_ops().back();
}
else
{
continue;
}
three_prime_ends.push_back(make_pair(scaff.fpkm(), &scaff));
}
}
adjacency_list <vecS, vecS, undirectedS> G;
for (size_t i = 0; i < three_prime_ends.size(); ++i)
{
add_vertex(G);
}
for (size_t i = 0; i < three_prime_ends.size(); ++i)
{
Scaffold* scaff_i = three_prime_ends[i].second;
//assert (scaff_i);
const AugmentedCuffOp* scaff_i_exon_3 = NULL;
if (scaff_i->strand() == CUFF_REV)
{
scaff_i_exon_3 = &(scaff_i->augmented_ops().front());
}
else if (scaff_i->strand() == CUFF_FWD)
{
scaff_i_exon_3 = &(scaff_i->augmented_ops().back());
}
for (size_t j = i + 1; j < three_prime_ends.size(); ++j)
{
Scaffold* scaff_j = three_prime_ends[j].second;
if (scaff_i->strand() != scaff_j->strand())
continue;
const AugmentedCuffOp* scaff_j_exon_3 = NULL;
if (scaff_j->strand() == CUFF_REV)
{
scaff_j_exon_3 = &(scaff_j->augmented_ops().front());
}
else if (scaff_j->strand() == CUFF_FWD)
{
scaff_j_exon_3 = &(scaff_j->augmented_ops().back());
}
if (AugmentedCuffOp::overlap_in_genome(*scaff_j_exon_3, *scaff_i_exon_3) &&
AugmentedCuffOp::compatible(*scaff_j_exon_3, *scaff_i_exon_3, 0))
add_edge(i, j, G);
}
}
std::vector<int> component(num_vertices(G));
connected_components(G, &component[0]);
vector<vector<bool> > clusters(three_prime_ends.size(),
vector<bool>(three_prime_ends.size(), false));
//vector<vector<size_t> > cluster_indices(three_prime_ends.size());
vector<vector<pair<double, Scaffold*> > > grouped_scaffolds(three_prime_ends.size());
for (size_t i = 0; i < three_prime_ends.size(); ++i)
{
clusters[component[i]][i] = true;
grouped_scaffolds[component[i]].push_back(three_prime_ends[i]);
}
for (size_t i = 0; i < grouped_scaffolds.size(); ++i)
{
vector<pair<double, Scaffold*> >& group = grouped_scaffolds[i];
sort(group.begin(), group.end());
if (group.empty())
continue;
Scaffold* group_leader = NULL;
int trim_point = -1;
const AugmentedCuffOp* group_exon_3 = NULL;
vector<pair<double, Scaffold*> >::iterator l_itr = group.begin();
while (l_itr != group.end())
{
Scaffold* possible_leader = l_itr->second;
bool ok_clip_leader = true;
vector<pair<double, Scaffold*> >::iterator g_itr = group.begin();
const AugmentedCuffOp* l_exon_3 = NULL;
CuffStrand s = possible_leader->strand();
if (s != CUFF_STRAND_UNKNOWN)
{
if (s == CUFF_REV)
l_exon_3 = &(possible_leader->augmented_ops().front());
else
l_exon_3 = &(possible_leader->augmented_ops().back());
for (; g_itr != group.end(); ++g_itr)
{
const AugmentedCuffOp* g_exon_3 = NULL;
if (s == CUFF_REV)
{
// bad:
// leader
// follower
g_exon_3 = &(g_itr->second->augmented_ops().front());
if (g_exon_3->g_right() <= l_exon_3->g_left())
ok_clip_leader = false;
// for meta-assembly libraries, don't ever allow clipping, just extension
// bad:
// leader
// follower
if (library_type == "transfrags" &&
g_exon_3->g_left() < l_exon_3->g_left())
ok_clip_leader = false;
}
else
{
// bad:
// follower
// leader
g_exon_3 = &(g_itr->second->augmented_ops().back());
if (g_exon_3->g_left() >= l_exon_3->g_right())
ok_clip_leader = false;
// for meta-assembly libraries, don't ever allow clipping, just extension
// bad:
// leader
// follower
if (library_type == "transfrags" &&
g_exon_3->g_right() > l_exon_3->g_right())
ok_clip_leader = false;
}
}
}
else
{
ok_clip_leader = false;
}
if (ok_clip_leader)
{
if (s == CUFF_REV)
{
if (trim_point == -1)
trim_point = l_exon_3->g_left();
else if (l_exon_3->g_left() < trim_point)
ok_clip_leader = false;
}
else
{
if (trim_point == -1)
trim_point = l_exon_3->g_right();
else if (l_exon_3->g_right() > trim_point)
ok_clip_leader = false;
}
}
if (ok_clip_leader)
{
group_leader = possible_leader;
group_exon_3 = l_exon_3;
break;
}
++l_itr;
}
if (!group_leader || !group_exon_3)
continue;
for (size_t j = 0; j < group.size(); ++j)
{
const AugmentedCuffOp* exon_3 = NULL;
int end_diff = 0;
if (group_leader->strand() == CUFF_REV)
{
exon_3 = &(group[j].second->augmented_ops().front());
end_diff = group_exon_3->g_left() - exon_3->g_left();
}
else
{
exon_3 = &(group[j].second->augmented_ops().back());
end_diff = exon_3->g_right() - group_exon_3->g_right();
}
if (end_diff > 0)
{
// leader
// follower
group[j].second->trim_3(end_diff);
}
else if (end_diff < 0)
{
// leader
// follower
group[j].second->extend_3(-end_diff);
}
}
}
return;
}
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