Apply base quality score recalibration on Spark
Category Read Data Manipulation
Overview
Apply base quality score recalibration with Spark.
This tool performs the second pass in a two-stage process called Base Quality Score Recalibration (BQSR). Specifically, it recalibrates the base qualities of the input reads based on the recalibration table produced by the BaseRecalibrator tool, and outputs a recalibrated BAM or CRAM file.
See Tutorial#10060 for an example of how to set up and run a Spark tool on a cloud Spark cluster.
Usage examples
gatk ApplyBQSRSpark \ -I gs://my-gcs-bucket/input.bam \ -bqsr gs://my-gcs-bucket/recalibration.table \ -O gs://my-gcs-bucket/output.bam \ -- \ --sparkRunner GCS \ --cluster my-dataproc-cluster
To additionally bin base qualities:
gatk ApplyBQSRSpark \ -I gs://my-gcs-bucket/input.bam \ -bqsr gs://my-gcs-bucket/recalibration.table \ -SQQ 10 -SQQ 20 -SQQ 30 -SQQ 40 \ -O gs://my-gcs-bucket/output.bam \ -- \ --sparkRunner GCS \ --cluster my-dataproc-cluster
ApplyBQSRSpark specific arguments
This table summarizes the command-line arguments that are specific to this tool. For more details on each argument, see the list further down below the table or click on an argument name to jump directly to that entry in the list.
Argument name(s) | Default value | Summary | |
---|---|---|---|
Required Arguments | |||
--bqsr-recal-file -bqsr |
null | Input covariates table file for base quality score recalibration | |
--input -I |
[] | BAM/SAM/CRAM file containing reads | |
--output -O |
null | the output bam | |
Optional Tool Arguments | |||
--arguments_file |
[] | read one or more arguments files and add them to the command line | |
--bam-partition-size |
0 | maximum number of bytes to read from a file into each partition of reads. Setting this higher will result in fewer partitions. Note that this will not be equal to the size of the partition in memory. Defaults to 0, which uses the default split size (determined by the Hadoop input format, typically the size of one HDFS block). | |
--conf |
[] | spark properties to set on the spark context in the format = | |
--disable-sequence-dictionary-validation |
false | If specified, do not check the sequence dictionaries from our inputs for compatibility. Use at your own risk! | |
--emit-original-quals |
false | Emit original base qualities under the OQ tag | |
--gcs-max-retries -gcs-retries |
20 | If the GCS bucket channel errors out, how many times it will attempt to re-initiate the connection | |
--global-qscore-prior |
-1.0 | Global Qscore Bayesian prior to use for BQSR | |
--help -h |
false | display the help message | |
--interval-merging-rule -imr |
ALL | Interval merging rule for abutting intervals | |
--intervals -L |
[] | One or more genomic intervals over which to operate | |
--num-reducers |
0 | For tools that shuffle data or write an output, sets the number of reducers. Defaults to 0, which gives one partition per 10MB of input. | |
--preserve-qscores-less-than |
6 | Don't recalibrate bases with quality scores less than this threshold | |
--program-name |
null | Name of the program running | |
--quantize-quals |
0 | Quantize quality scores to a given number of levels | |
--reference -R |
null | Reference sequence | |
--sharded-output |
false | For tools that write an output, write the output in multiple pieces (shards) | |
--spark-master |
local[*] | URL of the Spark Master to submit jobs to when using the Spark pipeline runner. | |
--use-original-qualities -OQ |
false | Use the base quality scores from the OQ tag | |
--version |
false | display the version number for this tool | |
Optional Common Arguments | |||
--disable-read-filter -DF |
[] | Read filters to be disabled before analysis | |
--disable-tool-default-read-filters |
false | Disable all tool default read filters | |
--exclude-intervals -XL |
[] | One or more genomic intervals to exclude from processing | |
--gatk-config-file |
null | A configuration file to use with the GATK. | |
--interval-exclusion-padding -ixp |
0 | Amount of padding (in bp) to add to each interval you are excluding. | |
--interval-padding -ip |
0 | Amount of padding (in bp) to add to each interval you are including. | |
--interval-set-rule -isr |
UNION | Set merging approach to use for combining interval inputs | |
--QUIET |
false | Whether to suppress job-summary info on System.err. | |
--read-filter -RF |
[] | Read filters to be applied before analysis | |
--read-index |
[] | Indices to use for the read inputs. If specified, an index must be provided for every read input and in the same order as the read inputs. If this argument is not specified, the path to the index for each input will be inferred automatically. | |
--read-validation-stringency -VS |
SILENT | Validation stringency for all SAM/BAM/CRAM/SRA files read by this program. The default stringency value SILENT can improve performance when processing a BAM file in which variable-length data (read, qualities, tags) do not otherwise need to be decoded. | |
--TMP_DIR |
[] | Undocumented option | |
--use-jdk-deflater -jdk-deflater |
false | Whether to use the JdkDeflater (as opposed to IntelDeflater) | |
--use-jdk-inflater -jdk-inflater |
false | Whether to use the JdkInflater (as opposed to IntelInflater) | |
--verbosity |
INFO | Control verbosity of logging. | |
Advanced Arguments | |||
--round-down-quantized |
false | Round quals down to nearest quantized qual | |
--showHidden |
false | display hidden arguments | |
--static-quantized-quals |
[] | Use static quantized quality scores to a given number of levels (with -bqsr) |
Argument details
Arguments in this list are specific to this tool. Keep in mind that other arguments are available that are shared with other tools (e.g. command-line GATK arguments); see Inherited arguments above.
--arguments_file / NA
read one or more arguments files and add them to the command line
List[File] []
--bam-partition-size / NA
maximum number of bytes to read from a file into each partition of reads. Setting this higher will result in fewer partitions. Note that this will not be equal to the size of the partition in memory. Defaults to 0, which uses the default split size (determined by the Hadoop input format, typically the size of one HDFS block).
long 0 [ [ -∞ ∞ ] ]
--bqsr-recal-file / -bqsr
Input covariates table file for base quality score recalibration
Enables recalibration of base qualities.
The covariates tables are produced by the BaseRecalibrator tool.
Please be aware that you should only run recalibration with the covariates file created on the same input bam(s).
R String null
--conf / -conf
spark properties to set on the spark context in the format =
List[String] []
--disable-read-filter / -DF
Read filters to be disabled before analysis
List[String] []
--disable-sequence-dictionary-validation / -disable-sequence-dictionary-validation
If specified, do not check the sequence dictionaries from our inputs for compatibility. Use at your own risk!
boolean false
--disable-tool-default-read-filters / -disable-tool-default-read-filters
Disable all tool default read filters
boolean false
--emit-original-quals / NA
Emit original base qualities under the OQ tag
The tool is capable of writing out the original quality scores of each read in the recalibrated output file
under the "OQ" tag. By default, this behavior is disabled because emitting original qualities results in a
significant increase of the file size. Use this flag to turn on emission of original qualities.
boolean false
--exclude-intervals / -XL
One or more genomic intervals to exclude from processing
Use this argument to exclude certain parts of the genome from the analysis (like -L, but the opposite).
This argument can be specified multiple times. You can use samtools-style intervals either explicitly on the
command line (e.g. -XL 1 or -XL 1:100-200) or by loading in a file containing a list of intervals
(e.g. -XL myFile.intervals).
List[String] []
--gatk-config-file / NA
A configuration file to use with the GATK.
String null
--gcs-max-retries / -gcs-retries
If the GCS bucket channel errors out, how many times it will attempt to re-initiate the connection
int 20 [ [ -∞ ∞ ] ]
--global-qscore-prior / NA
Global Qscore Bayesian prior to use for BQSR
If specified, the value of this argument will be used as a flat prior for all mismatching quality scores instead
of the reported quality score (assigned by the sequencer).
double -1.0 [ [ -∞ ∞ ] ]
--help / -h
display the help message
boolean false
--input / -I
BAM/SAM/CRAM file containing reads
R List[String] []
--interval-exclusion-padding / -ixp
Amount of padding (in bp) to add to each interval you are excluding.
Use this to add padding to the intervals specified using -XL. For example, '-XL 1:100' with a
padding value of 20 would turn into '-XL 1:80-120'. This is typically used to add padding around targets when
analyzing exomes.
int 0 [ [ -∞ ∞ ] ]
--interval-merging-rule / -imr
Interval merging rule for abutting intervals
By default, the program merges abutting intervals (i.e. intervals that are directly side-by-side but do not
actually overlap) into a single continuous interval. However you can change this behavior if you want them to be
treated as separate intervals instead.
The --interval-merging-rule argument is an enumerated type (IntervalMergingRule), which can have one of the following values:
- ALL
- OVERLAPPING_ONLY
IntervalMergingRule ALL
--interval-padding / -ip
Amount of padding (in bp) to add to each interval you are including.
Use this to add padding to the intervals specified using -L. For example, '-L 1:100' with a
padding value of 20 would turn into '-L 1:80-120'. This is typically used to add padding around targets when
analyzing exomes.
int 0 [ [ -∞ ∞ ] ]
--interval-set-rule / -isr
Set merging approach to use for combining interval inputs
By default, the program will take the UNION of all intervals specified using -L and/or -XL. However, you can
change this setting for -L, for example if you want to take the INTERSECTION of the sets instead. E.g. to
perform the analysis only on chromosome 1 exomes, you could specify -L exomes.intervals -L 1 --interval-set-rule
INTERSECTION. However, it is not possible to modify the merging approach for intervals passed using -XL (they will
always be merged using UNION).
Note that if you specify both -L and -XL, the -XL interval set will be subtracted from the -L interval set.
The --interval-set-rule argument is an enumerated type (IntervalSetRule), which can have one of the following values:
- UNION
- Take the union of all intervals
- INTERSECTION
- Take the intersection of intervals (the subset that overlaps all intervals specified)
IntervalSetRule UNION
--intervals / -L
One or more genomic intervals over which to operate
List[String] []
--num-reducers / NA
For tools that shuffle data or write an output, sets the number of reducers. Defaults to 0, which gives one partition per 10MB of input.
int 0 [ [ -∞ ∞ ] ]
--output / -O
the output bam
R String null
--preserve-qscores-less-than / NA
Don't recalibrate bases with quality scores less than this threshold
This flag tells GATK not to modify quality scores less than this value. Instead they will be written out
unmodified in the recalibrated BAM file. In general it's unsafe to change qualities scores below < 6, since
base callers use these values to indicate random or bad bases. For example, Illumina writes Q2 bases when the
machine has really gone wrong. This would be fine in and of itself, but when you select a subset of these reads
based on their ability to align to the reference and their dinucleotide effect, your Q2 bin can be elevated to
Q8 or Q10, leading to issues downstream.
int 6 [ [ -∞ ∞ ] ]
--program-name / NA
Name of the program running
String null
--quantize-quals / NA
Quantize quality scores to a given number of levels
Turns on the base quantization module. It requires a recalibration report.
A value of 0 here means "do not quantize".
Any value greater than zero will be used to recalculate the quantization using that many levels.
Negative values mean that we should quantize using the recalibration report's quantization level.
Exclusion: This argument cannot be used at the same time as static-quantized-quals, round-down-quantized
.
int 0 [ [ -∞ ∞ ] ]
--QUIET / NA
Whether to suppress job-summary info on System.err.
Boolean false
--read-filter / -RF
Read filters to be applied before analysis
List[String] []
--read-index / -read-index
Indices to use for the read inputs. If specified, an index must be provided for every read input and in the same order as the read inputs. If this argument is not specified, the path to the index for each input will be inferred automatically.
List[String] []
--read-validation-stringency / -VS
Validation stringency for all SAM/BAM/CRAM/SRA files read by this program. The default stringency value SILENT can improve performance when processing a BAM file in which variable-length data (read, qualities, tags) do not otherwise need to be decoded.
The --read-validation-stringency argument is an enumerated type (ValidationStringency), which can have one of the following values:
- STRICT
- LENIENT
- SILENT
ValidationStringency SILENT
--reference / -R
Reference sequence
String null
--round-down-quantized / NA
Round quals down to nearest quantized qual
Round down quantized only works with the static_quantized_quals option, and should not be used with
the dynamic binning option provided by quantize_quals. When roundDown = false, rounding is done in
probability space to the nearest bin. When roundDown = true, the value is rounded to the nearest bin
that is smaller than the current bin.
Exclusion: This argument cannot be used at the same time as quantize-quals
.
boolean false
--sharded-output / NA
For tools that write an output, write the output in multiple pieces (shards)
boolean false
--showHidden / -showHidden
display hidden arguments
boolean false
--spark-master / NA
URL of the Spark Master to submit jobs to when using the Spark pipeline runner.
String local[*]
--static-quantized-quals / NA
Use static quantized quality scores to a given number of levels (with -bqsr)
Static quantized quals are entirely separate from the quantize_qual option which uses dynamic binning.
The two types of binning should not be used together.
Exclusion: This argument cannot be used at the same time as quantize-quals
.
List[Integer] []
--TMP_DIR / NA
Undocumented option
List[File] []
--use-jdk-deflater / -jdk-deflater
Whether to use the JdkDeflater (as opposed to IntelDeflater)
boolean false
--use-jdk-inflater / -jdk-inflater
Whether to use the JdkInflater (as opposed to IntelInflater)
boolean false
--use-original-qualities / -OQ
Use the base quality scores from the OQ tag
This flag tells GATK to use the original base qualities (that were in the data before BQSR/recalibration) which
are stored in the OQ tag, if they are present, rather than use the post-recalibration quality scores. If no OQ
tag is present for a read, the standard quality score will be used.
Boolean false
--verbosity / -verbosity
Control verbosity of logging.
The --verbosity argument is an enumerated type (LogLevel), which can have one of the following values:
- ERROR
- WARNING
- INFO
- DEBUG
LogLevel INFO
--version / NA
display the version number for this tool
boolean false
GATK version 4.0.2.0 built at 02-13-2019 02:13:49.
0 comments
Please sign in to leave a comment.