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package net.sf.basedb.reggie.plugins.cmd; |
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import java.util.Date; |
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import java.util.List; |
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import java.util.regex.Matcher; |
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import java.util.regex.Pattern; |
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|
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import net.sf.basedb.core.BioPlate; |
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import net.sf.basedb.core.BioWell; |
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import net.sf.basedb.core.DbControl; |
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import net.sf.basedb.core.ItemQuery; |
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import net.sf.basedb.core.Protocol; |
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import net.sf.basedb.core.Tag; |
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import net.sf.basedb.core.data.PlateCoordinate; |
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import net.sf.basedb.core.query.Annotations; |
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import net.sf.basedb.core.query.Expressions; |
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import net.sf.basedb.core.query.Hql; |
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import net.sf.basedb.core.query.Restrictions; |
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import net.sf.basedb.reggie.Reggie; |
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import net.sf.basedb.reggie.dao.Annotationtype; |
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import net.sf.basedb.reggie.dao.BioplateType; |
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import net.sf.basedb.reggie.dao.Pipeline; |
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import net.sf.basedb.reggie.dao.Subtype; |
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import net.sf.basedb.reggie.plugins.cmd.ScanBIdRef.SampleIdType; |
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import net.sf.basedb.util.Coordinate; |
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/** |
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Holds all information about a library. Validation will |
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be done at construction and errors are reported to |
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the JsonSection. Check the 'valid' flag before using |
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the information. |
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|
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@since 4.32 |
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*/ |
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public class LibraryInfo |
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{ |
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|
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public String plateId; |
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public PlateCoordinate well; |
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|
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public Date libDate; |
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public Integer libSize; |
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public Float libMolarity; |
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public Float qubitConc; |
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public Float quantItConc; |
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public Float usedQuantityFromRna_ng; |
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public String operator; |
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|
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public BarcodeInfo barcodeInfo; |
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public Protocol protocol; |
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public BioPlate libPlate; |
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public BioplateType libPlateType = BioplateType.EXTERNAL_LIBRARY; |
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|
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public boolean valid; |
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|
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public LibraryInfo(JsonSection section, RnaInfo rna, FastqInfo fastqInfo, MainInfo main, ImportContext ctx) |
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{ |
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if (section != null) |
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{ |
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plateId = section.getRequiredEntry("Plate name", PatternValidator.CMD_ID); |
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ValidPlateCoordinateWithType plateWell = section.getRequiredEntry("Plate well", PlateOrStripValidator.INSTANCE); |
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if (plateWell != null) |
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{ |
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well = plateWell.plateCoordinate; |
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libPlateType = plateWell.plateType; |
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} |
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|
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if (plateId != null && well != null && ctx != null) |
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{ |
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JsonSection duplicate = ctx.add("LIB:"+plateId+":"+well, section); |
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if (duplicate != null) |
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{ |
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String coordinate = Coordinate.numericToAlpha(well.getRow()+1)+(well.getColumn()+1); |
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String msg = "Plate well ["+coordinate+"; ExternalRef="+plateId+"] duplicated in file: "; |
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section.addErrorMessage(msg+duplicate.getFile().getName()); |
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duplicate.addErrorMessage(msg+section.getFile().getName()); |
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} |
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} |
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|
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if (plateId != null) libPlate = findExistingLibPlate(plateId, libPlateType, section); |
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if (libPlate != null && well != null) |
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{ |
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verifyEmptyPosition(libPlate, well, section); |
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} |
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|
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libDate = section.getRequiredEntry("Date", DateValidator.YYYY_MM_DD.warnIfFutureOrOlder(rna!=null?rna.qiacubeDate:null, main.refDate)); |
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libSize = section.getRequiredEntry("Library size", IntValidator.POSITIVE.warnIf(150, 500)); |
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libMolarity = section.getRequiredEntry("Library molarity", FloatValidator.POSITIVE); |
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qubitConc = section.getOptionalEntry("Qubit concentration (ng/ul)", NullValidator.allowNull(FloatValidator.POSITIVE)); |
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quantItConc = section.getOptionalEntry("QuantIT concentration (ng/ul)", NullValidator.allowNull(FloatValidator.POSITIVE)); |
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if (qubitConc == null && quantItConc == null) |
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{ |
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section.addErrorMessage("Both Qubit and QuantIT concentrations are missing in "+section.getName()+" section"); |
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} |
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|
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operator = section.getOptionalEntry("Operator", null); |
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|
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barcodeInfo = section.getRequiredEntry("Barcode", BarcodeValidator.INSTANCE); |
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|
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if (barcodeInfo != null && fastqInfo != null && fastqInfo.barcodeSequences != null) |
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{ |
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String libSeq = barcodeInfo.seq1+"+"+BarcodeUtil.reverseComplement(barcodeInfo.seq2); |
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if (!BarcodeUtil.matches(fastqInfo.barcodeSequences, libSeq, 4, 2)) |
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{ |
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section.addErrorMessage("Library.Barcode doesn't match FASTQ header: "+libSeq+" != "+fastqInfo.barcodeSequences); |
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} |
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} |
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|
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protocol = section.getRequiredEntry("Protocol", ProtocolValidator.LIB_PROTOCOL); |
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|
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SampleIdType idType = main.idRef != null ? main.idRef.idType : null; |
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if (idType != SampleIdType.PRENORMALISED_RNA) |
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{ |
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if (rna != null && rna.originalQuantity_ng != null && rna.remainingQuantity_ng != null) |
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{ |
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usedQuantityFromRna_ng = rna.originalQuantity_ng-rna.remainingQuantity_ng; |
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} |
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} |
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} |
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valid = section != null && !section.hasError(); |
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} |
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|
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private BioPlate findExistingLibPlate(String plateId, BioplateType plateType, JsonSection section) |
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{ |
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BioPlate libPlate = null; |
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DbControl dc = section.getFile().dc(); |
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ItemQuery<BioPlate> query = BioPlate.getQuery(); |
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query.setIncludes(Reggie.INCLUDE_IN_CURRENT_PROJECT); |
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if (plateType != null) |
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{ |
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plateType.addFilter(dc, query, false); |
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} |
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query.join(Annotations.innerJoin(null, Annotationtype.EXTERNAL_REF.load(dc), "eref")); |
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query.restrict(Restrictions.eq(Hql.alias("eref"), Expressions.string(plateId))); |
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List<BioPlate> list = query.list(dc); |
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if (list.size() > 1) |
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{ |
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section.addErrorMessage("Found "+list.size()+" library plates with ExternalRef="+plateId); |
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} |
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else if (list.size() == 1) |
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{ |
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libPlate = list.get(0); |
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} |
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return libPlate; |
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} |
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|
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/** |
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Verify that the given position on the libplate is empty. |
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*/ |
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private void verifyEmptyPosition(BioPlate libPlate, PlateCoordinate pos, JsonSection section) |
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{ |
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String coordinate = Coordinate.numericToAlpha(pos.getRow()+1)+(pos.getColumn()+1); |
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BioWell well = libPlate.getBioWell(pos); |
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if (well == null) |
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{ |
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section.addErrorMessage("Well not found: "+libPlate.getName()+"["+ coordinate+"; ExternalRef="+plateId+"]"); |
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} |
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else if (!well.isEmpty()) |
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{ |
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section.addErrorMessage("Well is not empty: "+libPlate.getName()+"["+ coordinate+"; ExternalRef="+plateId+"]"); |
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} |
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} |
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|
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|
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/** |
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Find a barcode by looking at the UDI part of the value from the JSON file. |
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The entry in the JSON is typically similar to one of: |
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* 04A UDI0025 (ACTAAGAT-AACCGCGG) |
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* 03B IDT10_UDI_Adapter2018 (ATCTGGCCAT-CCGCTGTCTA) |
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If a barcode is found we also check the Barcode sequences and issue |
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a warning if they don't match. |
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*/ |
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static class BarcodeValidator |
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implements ValueValidator<String, BarcodeInfo> |
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{ |
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|
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static final BarcodeValidator INSTANCE = new BarcodeValidator(); |
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|
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@Override |
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public Class<String> getExpectedClass() |
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{ |
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return String.class; |
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} |
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|
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@Override |
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public BarcodeInfo isValid(DbControl dc, String value, JsonSection section, String entryKey) |
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{ |
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Pattern p = Pattern.compile("\\w+\\s+((?:UDI\\d+)|(?:IDT10\\w+))\\s+\\(([ACGT]+)-([ACGT]+)\\)"); |
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Matcher m = p.matcher(value); |
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if (!m.matches()) |
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{ |
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section.addErrorMessage("Invalid barcode in JSON: "+entryKey+"="+value+" (expected to find 'UDI[0-9]+' or 'IDT10...')"); |
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return null; |
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} |
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|
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BarcodeInfo bc = new BarcodeInfo(); |
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bc.raw = value; |
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bc.name = m.group(1); |
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bc.barcodeSet = bc.name.startsWith("IDT") ? "IDT10_UDI" : "TruSeqUniqueDual"; |
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bc.seq1 = m.group(2); |
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bc.seq2 = m.group(3); |
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|
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ItemQuery<Tag> query = Tag.getQuery(); |
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query.setIncludes(Reggie.INCLUDE_IN_CURRENT_PROJECT); |
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Subtype.BARCODE.addFilter(dc, query); |
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Pipeline.RNA_SEQ.addFilter(dc, query); |
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query.restrict(Restrictions.eq(Hql.property("name"), Expressions.string(bc.name))); |
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List<Tag> list = query.list(dc); |
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if (list.size() > 1) |
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{ |
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section.addErrorMessage("Found "+list.size()+" barcodes: name="+bc.name); |
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return null; |
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} |
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else if (list.size() == 1) |
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{ |
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bc.tag = list.get(0); |
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String seq1 = (String)Annotationtype.BARCODE_SEQUENCE.getAnnotationValue(dc, bc.tag); |
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String seq2 = (String)Annotationtype.BARCODE_SEQUENCE_2.getAnnotationValue(dc, bc.tag); |
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if (!bc.seq1.equals(seq1) || !bc.seq2.equals(seq2)) |
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{ |
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section.addWarningMessage("Barcode sequence mismatch: "+entryKey+"="+value+" (expected "+seq1+"-"+seq2+")"); |
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} |
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} |
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return bc; |
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} |
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} |
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|
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/** |
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Barcode information from the JSON file, optionally |
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mapped to a Tag item in BASE. The 'barcodeSet' is |
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determined by name pattern: |
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UDI... -> TruSeqUniqueDual |
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IDT10... -> IDT10_UDI |
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*/ |
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public static class BarcodeInfo |
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{ |
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public String raw; |
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public String name; |
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public String seq1; |
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public String seq2; |
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public String barcodeSet; |
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public Tag tag; |
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} |
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|
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/** |
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Validator implementation that delegates to either STRIP_1x8 validator if |
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the coordinate is specified as 'digit:digit' or to PLATE_8x12 otherwise. |
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@since 4.33.4 |
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*/ |
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static class PlateOrStripValidator |
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implements ValueValidator<String, ValidPlateCoordinateWithType> |
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{ |
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static final PlateOrStripValidator INSTANCE = new PlateOrStripValidator(); |
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|
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@Override |
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public Class<String> getExpectedClass() |
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{ |
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return String.class; |
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04 Nov 21 |
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} |
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|
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@Override |
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public ValidPlateCoordinateWithType isValid(DbControl dc, String value, JsonSection section, String entryKey) |
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{ |
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ValidPlateCoordinateWithType result = new ValidPlateCoordinateWithType(); |
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PlateWellValidator validator; |
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if (value.matches("\\d\\:\\d")) |
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{ |
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validator = PlateWellValidator.STRIP_1x8; |
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result.plateType = BioplateType.EXTERNAL_LIBRARY_STRIP; |
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} |
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else |
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{ |
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validator = PlateWellValidator.PLATE_8x12; |
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result.plateType = BioplateType.EXTERNAL_LIBRARY; |
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} |
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result.plateCoordinate = validator.isValid(dc, value, section, entryKey); |
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return result; |
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} |
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} |
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04 Nov 21 |
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|
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04 Nov 21 |
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/** |
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04 Nov 21 |
nicklas |
Holds the validated plate type and coordinate. |
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04 Nov 21 |
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*/ |
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04 Nov 21 |
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static class ValidPlateCoordinateWithType |
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{ |
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PlateCoordinate plateCoordinate; |
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BioplateType plateType; |
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} |
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} |