Bc generalize attach barcodes

setup.py

@@ -37,6 +37,7 @@
     ],
     entry_points={
             'console_scripts': [
+                'AttachBarcodes = sctools.platform:BarcodePlatform.attach_barcodes',
                 'Attach10xBarcodes = sctools.platform:TenXV2.attach_barcodes',
                 'SplitBam = sctools.platform:GenericPlatform.split_bam',
                 'CalculateGeneMetrics = sctools.platform:GenericPlatform.calculate_gene_metrics',

src/sctools/platform.py

@@ -629,3 +629,318 @@ def attach_barcodes(cls, args=None):
         cls._tag_bamfile(args.u2, args.output_bamfile, tag_generators)
 
         return 0
+
+
+class BarcodePlatform(GenericPlatform):
+    """Command Line Interface for extracting and attaching barcodes with specified positions
+     generalizing TenXV2 attach barcodes
+
+    Sample, cell and/or molecule barcodes can be extracted and attached to an unmapped bam when the
+    corresponding barcode's start position and and length are provided. The sample barcode is extracted
+    from the index i7 fastq file and the cell and molecule barcode are extracted from the r1 fastq file
+
+    This class defines several methods that are created as CLI tools when sctools is installed
+    (see setup.py)
+
+    Attributes
+    ----------
+    cell_barcode : fastq.EmbeddedBarcode
+        A data class that defines the start and end position of the cell barcode and the tags to
+        assign the sequence and quality of the cell barcode
+    molecule_barcode : fastq.EmbeddedBarcode
+        A data class that defines the start and end position of the molecule barcode and the tags
+        to assign the sequence and quality of the molecule barcode
+    sample_barcode : fastq.EmbeddedBarcode
+        A data class that defines the start and end position of the sample barcode and the tags
+        to assign the sequence and quality of the sample barcode
+
+    Methods
+    -------
+    attach_barcodes()
+        Attach barcodes from the forward (r1) and optionally index (i1) fastq files to the reverse
+        (r2) bam file
+
+    """
+    cell_barcode = None
+    molecule_barcode = None
+    sample_barcode = None
+
+    @classmethod
+    def _validate_barcode_args(cls, args):
+        """Validates that the barcode start position is greater than 0
+
+        Parameters
+        ----------
+        args : object
+            arguments list, The default value of None, when passed to `parser.parse_args`
+            causes the parser to read `sys.argv`
+
+        Returns
+        -------
+        args : object
+            return arguments list if valid
+
+        """
+        # check that if a barcode start position is provided, its length is also (and vice versa)
+        cls._validate_barcode_length_and_position(args.cell_barcode_start_pos, args.cell_barcode_length)
+        cls._validate_barcode_length_and_position(args.molecule_barcode_start_pos, args.molecule_barcode_length)
+        cls._validate_barcode_length_and_position(args.sample_barcode_start_pos, args.sample_barcode_length)
+
+        # check that an index fastq is provided sample barcode length and position are given
+        if args.i1 is None and args.sample_barcode_length:
+            raise argparse.ArgumentError('An i7 index fastq file must be given to attach a sample barcode')
+
+        # check that cell and molecule barcodes don't overlap
+        if args.cell_barcode_length and args.molecule_barcode_length:
+            cls._validate_barcode_input(args.molecule_barcode_start_pos,
+                                        args.cell_barcode_start_pos + args.cell_barcode_length)
+
+        return args
+
+    @classmethod
+    def _validate_barcode_length_and_position(cls, barcode_start_position, barcode_length):
+        """Checks that either that both barcode length and position are given or that neither are given as arguments
+
+        Parameters
+        ----------
+        barcode_start_position : int
+            the user defined start position (base pairs) of the barcode
+
+        barcode_length : int
+            the user defined length (base pairs) of the barcode
+
+        Returns
+        -------
+        given_value : int
+            return given value if valid
+
+        """
+        barcode_start_pos_exists = bool(barcode_start_position) or (barcode_start_position == 0)
+        barcode_length_exists =  bool(barcode_length)
+        # (XOR boolean logic)
+        if (barcode_start_pos_exists != barcode_length_exists):
+            raise argparse.ArgumentError('Invalid position/length, both position and length must be provided by the user together')
+
+    @classmethod
+    def _validate_barcode_input(cls, given_value, min_value):
+        """Validates that the barcode input is greater than a min value
+
+        Parameters
+        ----------
+        given_value : int
+            the given value that must be greater than the min_value,
+            (barcode length or barcode starting position)
+
+        min_value : int
+            the min value that the given_value must be greater than
+
+        Returns
+        -------
+        given_value : int
+            return given value if valid
+
+        """
+        if given_value < min_value:
+            raise argparse.ArgumentTypeError('Invalid barcode length/position')
+        return given_value
+
+    @classmethod
+    def _validate_barcode_start_pos(cls, given_value):
+        """Validates that the barcode start position is greater than 0
+
+        Parameters
+        ----------
+        given_value : Union[int, str]
+            the given start position of the barcode to validate
+
+        Returns
+        -------
+        given_value : int
+            returns the start position if it is valid
+
+        """
+        return cls._validate_barcode_input(int(given_value), 0)
+
+    @classmethod
+    def _validate_barcode_length(cls, given_value):
+        """Validates that the barcode length  is greater than 1
+
+        Parameters
+        ----------
+        given_value : Union[int, str]
+            the given length of the barcode to validate
+
+        Returns
+        -------
+        given_value : int
+            returns the length if it is valid
+
+        """
+        return cls._validate_barcode_input(int(given_value), 1)
+
+    @classmethod
+    def _tag_bamfile(cls,
+                     input_bamfile_name: str,
+                     output_bamfile_name: str,
+                     tag_generators: Iterable[fastq.EmbeddedBarcodeGenerator]) -> None:
+        """Adds tags from fastq file(s) to a bam file.
+
+        Attaches tags extracted from fastq files by `tag_generators`, attaches them to records from
+        `input_bamfile_name`, and writes the result to `output_bamfile_name`
+
+        Parameters
+        ----------
+        input_bamfile_name : str
+            input bam
+        output_bamfile_name : str
+            output bam
+        tag_generators : Iterable[fastq.EmbeddedBarcodeGenerator]
+            Iterable of generators that yield barcodes from fastq files
+
+        """
+        bam_tagger = bam.Tagger(input_bamfile_name)
+        bam_tagger.tag(output_bamfile_name, tag_generators)
+
+    @classmethod
+    def _make_tag_generators(cls, r1, i1=None, whitelist=None) -> List[fastq.EmbeddedBarcodeGenerator]:
+        """Create tag generators from fastq files.
+
+        Tag generators are iterators that run over fastq records, they extract and yield all of the
+        barcodes embedded in each fastq record. This means extracting the cell, umi, and/or the sample barcode.
+
+        Parameters
+        ----------
+        r1 : str
+            forward fastq file, where possibly the cell and/or molecule barcode is found
+        i1 : str, optional
+            index fastq file, where the sample barcode is found
+        whitelist : str, optional
+            A file that contains a list of acceptable cell barcodes
+
+        Returns
+        -------
+        tag_generators : List[EmbeddedBarcodeGenerator]
+            EmbeddedBarcodeGenerators containing barcodes from the given fastq
+
+        """
+        tag_generators = []
+        barcode_args = {'fastq_files': r1}
+
+        if i1:
+            barcode_args['embedded_barcodes'] = [cls.sample_barcode]
+            tag_generators.append(fastq.EmbeddedBarcodeGenerator(**barcode_args))
+
+        if whitelist:
+            barcode_args['whitelist'] = whitelist
+            if cls.cell_barcode:
+                barcode_args['embedded_cell_barcode'] = cls.cell_barcode
+            if cls.molecule_barcode:
+                barcode_args['other_embedded_barcodes'] = cls.molecule_barcode
+            tag_generators.append(fastq.BarcodeGeneratorWithCorrectedCellBarcodes(**barcode_args))
+
+        else:
+            # for all the barcodes that have a length and starting position specified
+            barcode_args['embedded_barcodes'] = [barcode for barcode in [cls.cell_barcode, cls.molecule_barcode] if barcode]
+            tag_generators.append(fastq.EmbeddedBarcodeGenerator(**barcode_args))
+
+        return tag_generators
+
+    @classmethod
+    def attach_barcodes(cls, args=None):
+        """Command line entrypoint for attaching barcodes to a bamfile.
+
+        Parameters
+        ----------
+        args : Iterable[str], optional
+            arguments list, The default value of None, when passed to `parser.parse_args`
+            causes the parser to read `sys.argv`
+
+        Returns
+        -------
+        return_call : 0
+            return call if the program completes successfully
+
+        """
+        parser = argparse.ArgumentParser()
+        parser.add_argument('--r1',
+                            required=True,
+                            help='read 1 fastq file, where the cell and molecule barcode is found')
+        parser.add_argument('--u2',
+                            required=True,
+                            help='unaligned bam, can be converted from fastq read 2'
+                                 'using picard FastqToSam')
+        parser.add_argument('-o',
+                            '--output-bamfile',
+                            required=True,
+                            help='filename for tagged bam')
+        parser.add_argument('-w',
+                            '--whitelist',
+                            default=None,
+                            help='optional cell barcode whitelist. If provided, corrected barcodes '
+                                 'will also be output when barcodes are observed within 1ED of a '
+                                 'whitelisted barcode')
+        parser.add_argument('--i1',
+                            default=None,
+                            help='(optional) i7 index fastq file, where the sample barcode is found')
+        parser.add_argument('--sample-barcode-start-position',
+                            dest='sample_barcode_start_pos',
+                            default=None,
+                            help='the user defined start position (base pairs) of the sample barcode',
+                            type=cls._validate_barcode_start_pos)
+        parser.add_argument('--sample-barcode-length',
+                            dest='sample_barcode_length',
+                            default=None,
+                            help='the user defined length (base pairs) of the sample barcode',
+                            type=cls._validate_barcode_length)
+        parser.add_argument('--cell-barcode-start-position',
+                            dest='cell_barcode_start_pos',
+                            default=None,
+                            help='the user defined start position, in base pairs, of the cell barcode',
+                            type=cls._validate_barcode_start_pos)
+        parser.add_argument('--cell-barcode-length',
+                            dest='cell_barcode_length',
+                            default=None,
+                            help='the user defined length, in base pairs, of the cell barcode',
+                            type=cls._validate_barcode_length)
+        parser.add_argument('--molecule-barcode-start-position',
+                            dest='molecule_barcode_start_pos',
+                            default=None,
+                            help='the user defined start position, in base pairs, of the molecule barcode '
+                                 '(must be not overlap cell barcode if cell barcode is provided)',
+                            type=cls._validate_barcode_start_pos)
+        parser.add_argument('--molecule-barcode-length',
+                            dest='molecule_barcode_length',
+                            default=None,
+                            help='the user defined length, in base pairs, of the molecule barcode',
+                            type=cls._validate_barcode_length)
+
+        # parse and validate the args
+        if args:
+            args = parser.parse_args(args)
+        else:
+            args = parser.parse_args()
+        cls._validate_barcode_args(args)
+
+        # if the length and there for the start pos have been given as args
+        # get the appropriate barcodes
+        if args.cell_barcode_length:
+            cls.cell_barcode = fastq.EmbeddedBarcode(start=args.cell_barcode_start_pos,
+                                                     end=args.cell_barcode_start_pos + args.cell_barcode_length,
+                                                     quality_tag=consts.QUALITY_CELL_BARCODE_TAG_KEY,
+                                                     sequence_tag=consts.RAW_CELL_BARCODE_TAG_KEY)
+        if args.molecule_barcode_length:
+            cls.molecule_barcode = fastq.EmbeddedBarcode(start=args.molecule_barcode_start_pos,
+                                                         end=args.molecule_barcode_start_pos + args.molecule_barcode_length,
+                                                         quality_tag=consts.QUALITY_MOLECULE_BARCODE_TAG_KEY,
+                                                         sequence_tag=consts.RAW_MOLECULE_BARCODE_TAG_KEY)
+        if args.sample_barcode_length:
+            cls.sample_barcode = fastq.EmbeddedBarcode(start=args.sample_barcode_start_pos,
+                                                       end=args.sample_barcode_start_pos + args.sample_barcode_length,
+                                                       quality_tag=consts.QUALITY_SAMPLE_BARCODE_TAG_KEY,
+                                                       sequence_tag=consts.RAW_SAMPLE_BARCODE_TAG_KEY)
+
+        # make the tags and attach the barcodes
+        tag_generators = cls._make_tag_generators(args.r1, args.i1, args.whitelist)
+        cls._tag_bamfile(args.u2, args.output_bamfile, tag_generators)
+
+        return 0