Beispiel #1
0
    def run(self):
        """Run"""
        logging.debug("root_dir: {d}.".format(d=self.root_dir))
        logging.debug("nfl_fa: {f}.".format(f=self.nfl_fa))
        logging.debug("Total number of chunks: N={N}.".format(N=self.N))

        # Validate input files,
        (num_reads, reads_per_split, nfl_dir, splitted_fas_todo) = \
            self.validate_inputs()

        logging.info("Total number of reads is {n}.".format(n=num_reads))
        logging.info("Splitting nfl_fa into chunks each " +
                     "containing {n} reads.".format(n=reads_per_split))

        splitted_fas_done = splitFasta(input_fasta=real_ppath(self.nfl_fa),
                                       reads_per_split=reads_per_split,
                                       out_dir=nfl_dir,
                                       out_prefix="input.split")

        logging.info("Splitted files are: " + "\n".join(splitted_fas_done))
        for fa in splitted_fas_todo:
            if fa not in splitted_fas_done:
                logging.info("touching {f}".format(f=fa))
                touch(fa)
    def run(self):
        """Run"""
        logging.debug("root_dir: {d}.".format(d=self.root_dir))
        logging.debug("nfl_fa: {f}.".format(f=self.nfl_fa))
        logging.debug("Total number of chunks: N={N}.".format(N=self.N))

        # Validate input files,
        (num_reads, reads_per_split, nfl_dir, splitted_fas_todo) = self.validate_inputs()

        logging.info("Total number of reads is {n}.".format(n=num_reads))
        logging.info("Splitting nfl_fa into chunks each " + "containing {n} reads.".format(n=reads_per_split))

        splitted_fas_done = splitFasta(
            input_fasta=real_ppath(self.nfl_fa),
            reads_per_split=reads_per_split,
            out_dir=nfl_dir,
            out_prefix="input.split",
        )

        logging.info("Splitted files are: " + "\n".join(splitted_fas_done))
        for fa in splitted_fas_todo:
            if fa not in splitted_fas_done:
                logging.info("touching {f}".format(f=fa))
                touch(fa)
Beispiel #3
0
def build_uc_from_partial_daligner(input_fastq, ref_fasta, out_pickle,
                          ccs_fofn=None,
                          done_filename=None, use_finer_qv=False, cpus=24, no_qv_or_aln_checking=True):
    """
    Given an input_fasta file of non-full-length (partial) reads and
    (unpolished) consensus isoforms sequences in ref_fasta, align reads to
    consensus isoforms using BLASR, and then build up a mapping between
    consensus isoforms and reads (i.e., assign reads to isoforms).
    Finally, save
        {isoform_id: [read_ids],
         nohit: set(no_hit_read_ids)}
    to an output pickle file.

    ccs_fofn --- If None, assume no quality value is available,
    otherwise, use QV from ccs_fofn.
    """
    input_fastq = realpath(input_fastq)
    input_fasta = input_fastq[:input_fastq.rfind('.')] + '.fasta'
    ice_fq2fa(input_fastq, input_fasta)
    ref_fasta = realpath(ref_fasta)
    out_pickle = realpath(out_pickle)
    output_dir = os.path.dirname(out_pickle)

    daligner_sensitive_mode, _low, _high, _ignore5, _ignore3, _ece_min_len = get_daligner_sensitivity_setting(ref_fasta, is_fasta=True)

    # DB should always be already converted
    ref_obj = DazzIDHandler(ref_fasta, True)
    input_obj = DazzIDHandler(input_fasta, False)

    # ice_partial is already being called through qsub, so run everything local!
    runner = DalignerRunner(input_fasta, ref_fasta, is_FL=False, same_strand_only=False, \
                            query_converted=True, db_converted=True, query_made=False, \
                            db_made=True, use_sge=False, cpus=cpus, sge_opts=None)
    las_filenames, las_out_filenames = runner.runHPC(min_match_len=_low, output_dir=output_dir, sensitive_mode=daligner_sensitive_mode)

    if no_qv_or_aln_checking:
        # not using QVs or alignment checking!
        # this probqv is just a DUMMY to pass to daligner_against_ref, which won't be used
        logging.info("Not using QV for partial_uc. Loading dummy QV.")
        probqv = ProbFromModel(.01, .07, .06)
    else:
#        if ccs_fofn is None:
#            logging.info("Loading probability from model (0.01,0.07,0.06)")
#            probqv = ProbFromModel(.01, .07, .06)
#        else:
        start_t = time.time()
        probqv = ProbFromFastq(input_fastq)
        logging.info("Loading QVs from {fq} took {s} secs".format(fq=input_fastq, s=time.time()-start_t))
# --------- comment out below since we are just using FASTQ / BAM
#            if use_finer_qv:
#                probqv = ProbFromQV(input_fofn=ccs_fofn, fasta_filename=input_fasta)
#                logging.info("Loading QVs from {i} + {f} took {s} secs".format(f=ccs_fofn, i=input_fasta,\
#                    s=time.time()-start_t))
#            else:
#                input_fastq = input_fasta[:input_fasta.rfind('.')] + '.fastq'
#                logging.info("Converting {i} + {f} --> {fq}".format(i=input_fasta, f=ccs_fofn, fq=input_fastq))
#                ice_fa2fq(input_fasta, ccs_fofn, input_fastq)
#                probqv = ProbFromFastq(input_fastq)
#                logging.info("Loading QVs from {fq} took {s} secs".format(fq=input_fastq, s=time.time()-start_t))
#                print >> sys.stderr, "Loading QVs from {fq} took {s} secs".format(fq=input_fastq, s=time.time()-start_t)

    logging.info("Calling dalign_against_ref ...")

    partial_uc = {}  # Maps each isoform (cluster) id to a list of reads
    # which can map to the isoform
    seen = set()  # reads seen
    logging.info("Building uc from DALIGNER hits.")

    for las_out_filename in las_out_filenames:
        start_t = time.time()
        hitItems = dalign_against_ref(input_obj, ref_obj, las_out_filename,
                                 is_FL=False,
                                 sID_starts_with_c=True,
                                 qver_get_func=probqv.get_smoothed,
                                 qvmean_get_func=probqv.get_mean,
                                 ece_penalty=1,
                                 ece_min_len=_ece_min_len,
                                 same_strand_only=False,
                                 no_qv_or_aln_checking=no_qv_or_aln_checking,
                                 max_missed_start=_ignore5,
                                 max_missed_end=_ignore3)
        for h in hitItems:
            if h.ece_arr is not None:
                if h.cID not in partial_uc:
                    partial_uc[h.cID] = set()
                partial_uc[h.cID].add(h.qID)
                seen.add(h.qID)
        logging.info("processing {0} took {1} sec".format(las_out_filename, time.time()-start_t))
        print >> sys.stderr, "processing {0} took {1} sec".format(las_out_filename, time.time()-start_t)

    for k in partial_uc:
        partial_uc[k] = list(partial_uc[k])

    allhits = set(r.name.split()[0] for r in FastaReader(input_fasta))

    logging.info("Counting reads with no hit.")
    nohit = allhits.difference(seen)

    logging.info("Dumping uc to a pickle: {f}.".format(f=out_pickle))
    with open(out_pickle, 'w') as f:
        dump({'partial_uc': partial_uc, 'nohit': nohit}, f)

    done_filename = realpath(done_filename) if done_filename is not None \
        else out_pickle + '.DONE'
    logging.debug("Creating {f}.".format(f=done_filename))
    touch(done_filename)

    # remove all the .las and .las.out filenames
    for file in las_filenames:
        os.remove(file)
    for file in las_out_filenames:
        os.remove(file)
Beispiel #4
0
def build_uc_from_partial(input_fasta, ref_fasta, out_pickle,
                          sa_file=None, ccs_fofn=None,
                          done_filename=None, blasr_nproc=12, use_finer_qv=False):
    """
    Given an input_fasta file of non-full-length (partial) reads and
    (unpolished) consensus isoforms sequences in ref_fasta, align reads to
    consensus isoforms using BLASR, and then build up a mapping between
    consensus isoforms and reads (i.e., assign reads to isoforms).
    Finally, save
        {isoform_id: [read_ids],
         nohit: set(no_hit_read_ids)}
    to an output pickle file.

    ccs_fofn --- If None, assume no quality value is available,
    otherwise, use QV from ccs_fofn.
    blasr_nproc --- equivalent to blasr -nproc, number of CPUs to use
    """
    input_fasta = realpath(input_fasta)
    m5_file = input_fasta + ".blasr"
    out_pickle = realpath(out_pickle)
    if sa_file is None:
        if op.exists(input_fasta + ".sa"):
            sa_file = input_fasta + ".sa"

    cmd = "blasr {i} ".format(i=real_upath(input_fasta)) + \
          "{r} -bestn 5 ".format(r=real_upath(ref_fasta)) + \
          "-nproc {n} -m 5 ".format(n=blasr_nproc) + \
          "-maxScore -1000 -minPctIdentity 85 " + \
          "-out {o} ".format(o=real_upath(m5_file))
    if sa_file is not None and op.exists(sa_file):
        cmd += "-sa {sa}".format(sa=real_upath(sa_file))

    logging.info("CMD: {cmd}".format(cmd=cmd))
    _out, _code, _msg = backticks(cmd)
    if _code != 0:
        errMsg = "Command failed: {cmd}\n{e}".format(cmd=cmd, e=_msg)
        logging.error(errMsg)
        raise RuntimeError(errMsg)
    
    if ccs_fofn is None:
        logging.info("Loading probability from model (0.01,0.07,0.06)")
        probqv = ProbFromModel(.01, .07, .06)
    else:
        start_t = time.time()
        if use_finer_qv:
            logging.info("Loading QVs from {i} + {f} took {s} secs".format(f=ccs_fofn, i=input_fasta,\
                    s=time.time()-start_t))
            probqv = ProbFromQV(input_fofn=ccs_fofn, fasta_filename=input_fasta)
        else:
            input_fastq = input_fasta[:input_fasta.rfind('.')] + '.fastq'
            logging.info("Converting {i} + {f} --> {fq}".format(i=input_fasta, f=ccs_fofn, fq=input_fastq))
            ice_fa2fq(input_fasta, ccs_fofn, input_fastq)
            logging.info("Loading QVs from {fq} took {s} secs".format(fq=input_fastq, s=time.time()-start_t))
            probqv = ProbFromFastq(input_fastq)


    logging.info("Calling blasr_against_ref ...")
    hitItems = blasr_against_ref(output_filename=m5_file,
                                 is_FL=False,
                                 sID_starts_with_c=True,
                                 qvmean_get_func=probqv.get_mean,
                                 qver_get_func=probqv.get_smoothed,
                                 ece_penalty=1,
                                 ece_min_len=20,
                                 same_strand_only=False,
                                 max_missed_start=200,
                                 max_missed_end=50)

    partial_uc = {}  # Maps each isoform (cluster) id to a list of reads
    # which can map to the isoform
    seen = set()  # reads seen
    logging.info("Building uc from BLASR hits.")
    for h in hitItems:
        if h.ece_arr is not None:
            if h.cID not in partial_uc:
                partial_uc[h.cID] = set()
            partial_uc[h.cID].add(h.qID)
            seen.add(h.qID)

    for k in partial_uc:
        partial_uc[k] = list(partial_uc[k])

    allhits = set(r.name.split()[0] for r in FastaReader(input_fasta))

    logging.info("Counting reads with no hit.")
    nohit = allhits.difference(seen)

    logging.info("Dumping uc to a pickle: {f}.".format(f=out_pickle))
    with open(out_pickle, 'w') as f:
        dump({'partial_uc': partial_uc, 'nohit': nohit}, f)

    os.remove(m5_file)

    done_filename = realpath(done_filename) if done_filename is not None \
        else out_pickle + '.DONE'
    logging.debug("Creating {f}.".format(f=done_filename))
    touch(done_filename)
Beispiel #5
0
def build_uc_from_partial_daligner(input_fasta, ref_fasta, out_pickle,
                          ccs_fofn=None,
                          done_filename=None, use_finer_qv=False, cpus=24, no_qv_or_aln_checking=True):
    """
    Given an input_fasta file of non-full-length (partial) reads and
    (unpolished) consensus isoforms sequences in ref_fasta, align reads to
    consensus isoforms using BLASR, and then build up a mapping between
    consensus isoforms and reads (i.e., assign reads to isoforms).
    Finally, save
        {isoform_id: [read_ids],
         nohit: set(no_hit_read_ids)}
    to an output pickle file.

    ccs_fofn --- If None, assume no quality value is available,
    otherwise, use QV from ccs_fofn.
    """
    input_fasta = realpath(input_fasta)
    ref_fasta = realpath(ref_fasta)
    out_pickle = realpath(out_pickle)
    output_dir = os.path.dirname(out_pickle)

    daligner_sensitive_mode, _low, _high = get_daligner_sensitivity_setting(ref_fasta)

    # DB should always be already converted
    ref_obj = DazzIDHandler(ref_fasta, True)
    input_obj = DazzIDHandler(input_fasta, False)

    # ice_partial is already being called through qsub, so run everything local!
    runner = DalignerRunner(input_fasta, ref_fasta, is_FL=False, same_strand_only=False, \
                            query_converted=True, db_converted=True, query_made=False, \
                            db_made=True, use_sge=False, cpus=cpus, sge_opts=None)
    las_filenames, las_out_filenames = runner.runHPC(min_match_len=300, output_dir=output_dir, sensitive_mode=daligner_sensitive_mode)

    if no_qv_or_aln_checking:
        # not using QVs or alignment checking!
        # this probqv is just a DUMMY to pass to daligner_against_ref, which won't be used
        logging.info("Not using QV for partial_uc. Loading dummy QV.")
        probqv = ProbFromModel(.01, .07, .06)
    else:
        if ccs_fofn is None:
            logging.info("Loading probability from model (0.01,0.07,0.06)")
            probqv = ProbFromModel(.01, .07, .06)
        else:
            start_t = time.time()
            if use_finer_qv:
                probqv = ProbFromQV(input_fofn=ccs_fofn, fasta_filename=input_fasta)
                logging.info("Loading QVs from {i} + {f} took {s} secs".format(f=ccs_fofn, i=input_fasta,\
                    s=time.time()-start_t))
            else:
                input_fastq = input_fasta[:input_fasta.rfind('.')] + '.fastq'
                logging.info("Converting {i} + {f} --> {fq}".format(i=input_fasta, f=ccs_fofn, fq=input_fastq))
                ice_fa2fq(input_fasta, ccs_fofn, input_fastq)
                probqv = ProbFromFastq(input_fastq)
                logging.info("Loading QVs from {fq} took {s} secs".format(fq=input_fastq, s=time.time()-start_t))
                print >> sys.stderr, "Loading QVs from {fq} took {s} secs".format(fq=input_fastq, s=time.time()-start_t)

    logging.info("Calling dalign_against_ref ...")

    partial_uc = {}  # Maps each isoform (cluster) id to a list of reads
    # which can map to the isoform
    seen = set()  # reads seen
    logging.info("Building uc from DALIGNER hits.")

    for las_out_filename in las_out_filenames:
        start_t = time.time()
        hitItems = dalign_against_ref(input_obj, ref_obj, las_out_filename,
                                 is_FL=False,
                                 sID_starts_with_c=True,
                                 qver_get_func=probqv.get_smoothed,
                                 qvmean_get_func=probqv.get_mean,
                                 ece_penalty=1,
                                 ece_min_len=20,
                                 same_strand_only=False,
                                 no_qv_or_aln_checking=no_qv_or_aln_checking)
        for h in hitItems:
            if h.ece_arr is not None:
                if h.cID not in partial_uc:
                    partial_uc[h.cID] = set()
                partial_uc[h.cID].add(h.qID)
                seen.add(h.qID)
        logging.info("processing {0} took {1} sec".format(las_out_filename, time.time()-start_t))
        print >> sys.stderr, "processing {0} took {1} sec".format(las_out_filename, time.time()-start_t)

    for k in partial_uc:
        partial_uc[k] = list(partial_uc[k])

    allhits = set(r.name.split()[0] for r in FastaReader(input_fasta))

    logging.info("Counting reads with no hit.")
    nohit = allhits.difference(seen)

    logging.info("Dumping uc to a pickle: {f}.".format(f=out_pickle))
    with open(out_pickle, 'w') as f:
        dump({'partial_uc': partial_uc, 'nohit': nohit}, f)

    done_filename = realpath(done_filename) if done_filename is not None \
        else out_pickle + '.DONE'
    logging.debug("Creating {f}.".format(f=done_filename))
    touch(done_filename)

    # remove all the .las and .las.out filenames
    for file in las_filenames:
        os.remove(file)
    for file in las_out_filenames:
        os.remove(file)
Beispiel #6
0
def build_uc_from_partial(input_fasta, ref_fasta, out_pickle,
                          sa_file=None, ccs_fofn=None,
                          done_filename=None, blasr_nproc=12, use_finer_qv=False):
    """
    Given an input_fasta file of non-full-length (partial) reads and
    (unpolished) consensus isoforms sequences in ref_fasta, align reads to
    consensus isoforms using BLASR, and then build up a mapping between
    consensus isoforms and reads (i.e., assign reads to isoforms).
    Finally, save
        {isoform_id: [read_ids],
         nohit: set(no_hit_read_ids)}
    to an output pickle file.

    ccs_fofn --- If None, assume no quality value is available,
    otherwise, use QV from ccs_fofn.
    blasr_nproc --- equivalent to blasr -nproc, number of CPUs to use
    """
    input_fasta = realpath(input_fasta)
    m5_file = input_fasta + ".blasr"
    out_pickle = realpath(out_pickle)
    if sa_file is None:
        if op.exists(input_fasta + ".sa"):
            sa_file = input_fasta + ".sa"

    cmd = "blasr {i} ".format(i=real_upath(input_fasta)) + \
          "{r} -bestn 5 ".format(r=real_upath(ref_fasta)) + \
          "-nproc {n} -m 5 ".format(n=blasr_nproc) + \
          "-maxScore -1000 -minPctIdentity 85 " + \
          "-out {o} ".format(o=real_upath(m5_file))
    if sa_file is not None and op.exists(sa_file):
        cmd += "-sa {sa}".format(sa=real_upath(sa_file))

    logging.info("CMD: {cmd}".format(cmd=cmd))
    _out, _code, _msg = backticks(cmd)
    if _code != 0:
        errMsg = "Command failed: {cmd}\n{e}".format(cmd=cmd, e=_msg)
        logging.error(errMsg)
        raise RuntimeError(errMsg)
    
    if ccs_fofn is None:
        logging.info("Loading probability from model (0.01,0.07,0.06)")
        probqv = ProbFromModel(.01, .07, .06)
    else:
        start_t = time.time()
        if use_finer_qv:
            logging.info("Loading QVs from {i} + {f} took {s} secs".format(f=ccs_fofn, i=input_fasta,\
                    s=time.time()-start_t))
            probqv = ProbFromQV(input_fofn=ccs_fofn, fasta_filename=input_fasta)
        else:
            input_fastq = input_fasta[:input_fasta.rfind('.')] + '.fastq'
            logging.info("Converting {i} + {f} --> {fq}".format(i=input_fasta, f=ccs_fofn, fq=input_fastq))
            ice_fa2fq(input_fasta, ccs_fofn, input_fastq)
            logging.info("Loading QVs from {fq} took {s} secs".format(fq=input_fastq, s=time.time()-start_t))
            probqv = ProbFromFastq(input_fastq)


    logging.info("Calling blasr_against_ref ...")
    hitItems = blasr_against_ref(output_filename=m5_file,
                                 is_FL=False,
                                 sID_starts_with_c=True,
                                 qvmean_get_func=probqv.get_mean,
                                 qver_get_func=probqv.get_smoothed,
                                 ece_penalty=1,
                                 ece_min_len=10,
                                 same_strand_only=False)

    partial_uc = {}  # Maps each isoform (cluster) id to a list of reads
    # which can map to the isoform
    seen = set()  # reads seen
    logging.info("Building uc from BLASR hits.")
    for h in hitItems:
        if h.ece_arr is not None:
            if h.cID not in partial_uc:
                partial_uc[h.cID] = set()
            partial_uc[h.cID].add(h.qID)
            seen.add(h.qID)

    for k in partial_uc:
        partial_uc[k] = list(partial_uc[k])

    allhits = set(r.name.split()[0] for r in FastaReader(input_fasta))

    logging.info("Counting reads with no hit.")
    nohit = allhits.difference(seen)

    logging.info("Dumping uc to a pickle: {f}.".format(f=out_pickle))
    with open(out_pickle, 'w') as f:
        dump({'partial_uc': partial_uc, 'nohit': nohit}, f)

    os.remove(m5_file)

    done_filename = realpath(done_filename) if done_filename is not None \
        else out_pickle + '.DONE'
    logging.debug("Creating {f}.".format(f=done_filename))
    touch(done_filename)
Beispiel #7
0
def build_uc_from_partial(input_fasta,
                          ref_fasta,
                          out_pickle,
                          sa_file=None,
                          ccs_fofn=None,
                          done_filename=None,
                          blasr_nproc=12):
    """Align consensus isoforms in ref_fasta and reads in input_fasta,
    and save mappings between isoforms and reads to out_pickle.
    ccs_fofn --- If None, assume no quality value is available,
    otherwise, use QV from ccs_fofn.
    blasr_nproc --- equivalent to blasr -nproc, number of CPUs to use
    """
    input_fasta = realpath(input_fasta)
    m5_file = input_fasta + ".blasr"
    out_pickle = realpath(out_pickle)
    if sa_file is None:
        if op.exists(input_fasta + ".sa"):
            sa_file = input_fasta + ".sa"

    cmd = "blasr {i} ".format(i=input_fasta) + \
          "{r} -bestn 5 ".format(r=ref_fasta) + \
          "-nproc {n} -m 5 ".format(n=blasr_nproc) + \
          "-maxScore -1000 -minPctIdentity 85 -out {o} ".format(o=m5_file)
    if sa_file is not None and op.exists(sa_file):
        cmd += "-sa {sa}".format(sa=sa_file)

    logging.info("CMD: {cmd}".format(cmd=cmd))
    _out, _code, _msg = backticks(cmd)
    if _code != 0:
        errMsg = "Command failed: {cmd}\n{e}".format(cmd=cmd, e=_msg)
        logging.error(errMsg)
        raise RuntimeError(errMsg)

    if ccs_fofn is None:
        logging.info("Loading probability from model")
        probqv = ProbFromModel(.01, .07, .06)
    else:
        logging.info("Loading probability from QV in {f}".format(f=ccs_fofn))
        probqv = ProbFromQV(input_fofn=ccs_fofn, fasta_filename=input_fasta)

    logging.info("Calling blasr_against_ref ...")
    hitItems = blasr_against_ref(output_filename=m5_file,
                                 is_FL=False,
                                 sID_starts_with_c=True,
                                 qver_get_func=probqv.get_smoothed,
                                 ece_penalty=1,
                                 ece_min_len=10,
                                 same_strand_only=False)

    partial_uc = {}  # Maps each isoform (cluster) id to a list of reads
    # which can map to the isoform
    seen = set()  # reads seen
    logging.info("Building uc from BLASR hits.")
    for h in hitItems:
        if h.ece_arr is not None:
            if h.cID not in partial_uc:
                partial_uc[h.cID] = []
            partial_uc[h.cID].append(h.qID)
            seen.add(h.qID)

    allhits = set(r.name.split()[0] for r in FastaReader(input_fasta))

    logging.info("Counting reads with no hit.")
    nohit = allhits.difference(seen)

    logging.info("Dumping uc to a pickle: {f}.".format(f=out_pickle))
    with open(out_pickle, 'w') as f:
        dump({'partial_uc': partial_uc, 'nohit': nohit}, f)

    os.remove(m5_file)

    done_filename = realpath(done_filename) if done_filename is not None \
            else out_pickle + '.DONE'
    logging.debug("Creating {f}.".format(f=done_filename))
    touch(done_filename)
def build_uc_from_partial(
    input_fasta, ref_fasta, out_pickle, sa_file=None, ccs_fofn=None, done_filename=None, blasr_nproc=12
):
    """Align consensus isoforms in ref_fasta and reads in input_fasta,
    and save mappings between isoforms and reads to out_pickle.
    ccs_fofn --- If None, assume no quality value is available,
    otherwise, use QV from ccs_fofn.
    blasr_nproc --- equivalent to blasr -nproc, number of CPUs to use
    """
    input_fasta = realpath(input_fasta)
    m5_file = input_fasta + ".blasr"
    out_pickle = realpath(out_pickle)
    if sa_file is None:
        if op.exists(input_fasta + ".sa"):
            sa_file = input_fasta + ".sa"

    cmd = (
        "blasr {i} ".format(i=input_fasta)
        + "{r} -bestn 5 ".format(r=ref_fasta)
        + "-nproc {n} -m 5 ".format(n=blasr_nproc)
        + "-maxScore -1000 -minPctIdentity 85 -out {o} ".format(o=m5_file)
    )
    if sa_file is not None and op.exists(sa_file):
        cmd += "-sa {sa}".format(sa=sa_file)

    logging.info("CMD: {cmd}".format(cmd=cmd))
    _out, _code, _msg = backticks(cmd)
    if _code != 0:
        errMsg = "Command failed: {cmd}\n{e}".format(cmd=cmd, e=_msg)
        logging.error(errMsg)
        raise RuntimeError(errMsg)

    if ccs_fofn is None:
        logging.info("Loading probability from model")
        probqv = ProbFromModel(0.01, 0.07, 0.06)
    else:
        logging.info("Loading probability from QV in {f}".format(f=ccs_fofn))
        probqv = ProbFromQV(input_fofn=ccs_fofn, fasta_filename=input_fasta)

    logging.info("Calling blasr_against_ref ...")
    hitItems = blasr_against_ref(
        output_filename=m5_file,
        is_FL=False,
        sID_starts_with_c=True,
        qver_get_func=probqv.get_smoothed,
        ece_penalty=1,
        ece_min_len=10,
        same_strand_only=False,
    )

    partial_uc = {}  # Maps each isoform (cluster) id to a list of reads
    # which can map to the isoform
    seen = set()  # reads seen
    logging.info("Building uc from BLASR hits.")
    for h in hitItems:
        if h.ece_arr is not None:
            if h.cID not in partial_uc:
                partial_uc[h.cID] = []
            partial_uc[h.cID].append(h.qID)
            seen.add(h.qID)

    allhits = set(r.name.split()[0] for r in FastaReader(input_fasta))

    logging.info("Counting reads with no hit.")
    nohit = allhits.difference(seen)

    logging.info("Dumping uc to a pickle: {f}.".format(f=out_pickle))
    with open(out_pickle, "w") as f:
        dump({"partial_uc": partial_uc, "nohit": nohit}, f)

    os.remove(m5_file)

    done_filename = realpath(done_filename) if done_filename is not None else out_pickle + ".DONE"
    logging.debug("Creating {f}.".format(f=done_filename))
    touch(done_filename)