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package barcode;
import java.util.ArrayList;
import java.util.Collection;
import java.util.Collections;
import java.util.HashMap;
import java.util.LinkedHashMap;
import java.util.LinkedHashSet;
import java.util.Map.Entry;
import java.util.Set;
import fileIO.ByteFile;
import fileIO.ByteStreamWriter;
import hiseq.IlluminaHeaderParser2;
import shared.LineParser1;
import shared.LineParserS1;
import shared.Tools;
import stream.Read;
import structures.ByteBuilder;
import structures.LongList;
/**
* Writes legacy stats files when demultiplexing, for compatibility.
* These are not very informative and not recommended for use.
* @author BBushnell
*
*/
public class LegacyFileWriter {
public LegacyFileWriter(){}
// Top_Unknown_Barcodes.csv
// Lane,index,index2,# Reads,% of Unknown Barcodes,% of All Reads
// 7,AGGAGTTCTA,GGGGGGGGGG,3216744,0.017843,0.000839
// 7,TCGAATGATC,GGGGGGGGGG,2794420,0.015501,0.000729
// 7,CTTACCTGCT,GGGGGGGGGG,2709266,0.015028,0.000707
// (1000 lines)
long writeTopUnknownBarcodes(final Set<String> expectedSet,
HashMap<String, String> assignmentMap, final byte delimiter,
final String fname, final int lane, final int lines,
boolean overwrite) {
if(fname==null) {return 0;}
ArrayList<Barcode> list=new ArrayList<Barcode>(counts);
Collections.sort(list);
ByteStreamWriter bsw=ByteStreamWriter.makeBSW(fname, overwrite, false, true);
// if(delimiter<=0) {
// bsw.println("Lane,index,,# Reads,% of Unknown Barcodes,% of All Reads");
// }else {
bsw.println("Lane,index,index2,# Reads,% of Unknown Barcodes,% of All Reads");
// }
long total=0;
long expected=0;
for(Barcode bc : list) {
total+=bc.count();
if(assignmentMap.containsKey(bc.name)) {expected+=bc.count();}
}
final long unexpected=total-expected;
final float invTotal=1f/Tools.max(1, total);
final float invUnexpected=1f/Tools.max(1, unexpected);
// System.err.println("total="+total+", expected="+expected+", unexpected="+unexpected);
int printed=0;
ByteBuilder bb=new ByteBuilder();
for(Barcode bc : list) {
if(!assignmentMap.containsKey(bc.name)) {
if(printed>=lines) {break;}
printed++;
bb.clear().append(lane).comma();
bc.appendIndex(bb, delimiter, 1).comma();
// if(delimiter>0) {
bc.appendIndex(bb, delimiter, 2).comma();//It's OK if there isn't one
// }
// System.err.println("count="+bc.count());
bb.append(bc.count()).comma();
bb.append(bc.count()*invUnexpected, 6).comma();
bb.append(bc.count()*invTotal, 6).nl();
bsw.print(bb);
}
}
bsw.poison();
return unexpected;
}
//// Index_Hopping_Counts.csv
//// Lane,SampleID,index,index2,# Reads,% of Hopped Reads,% of All Reads
//// 7,529965,CCGTGCGGTC,CCCGATATGG,55663469,,0.014523
//// 7,529966,CACAGTTGTC,TAGGCAGGCT,52701428,,0.013750
//// 7,529967,CCTCTTCAGA,GATCCAGTAG,48686653,,0.012703
//// ...
//// 7,,CCGTGCGGTC,TAGGCAGGCT,3821,0.000132,0.000001
//// 7,,CCGTGCGGTC,GATCCAGTAG,5645,0.000196,0.000001
//// 7,,CCGTGCGGTC,GCCTGGTAAG,4099,0.000142,0.000001
//// (all valid then all invalid pairs)
void writeIndexHoppingCounts(final Set<String> expectedSet,
LinkedHashMap<String,String> sampleMap, final byte delimiter, final String fname,
final int lane, boolean overwrite) {
if(delimiter<=0) {return;}
LinkedHashSet<String> leftSet=new LinkedHashSet<String>();
LinkedHashSet<String> rightSet=new LinkedHashSet<String>();
LineParserS1 lp=new LineParserS1(delimiter);
for(String s : expectedSet) {
lp.set(s);
assert(lp.terms()==2) : "Wrong number of barcodes in '"+s+
"'; expected 2 for delimiter "+(int)delimiter;
leftSet.add(lp.parseString(0));
rightSet.add(lp.parseString(1));
}
// System.err.println(expectedSet.size());
final HashMap<String, Barcode> countMap=new HashMap<String, Barcode>(Tools.max(8, counts.size()));
float allMult=1f/Tools.max(1, (totalR1));
for(Barcode bc : counts) {countMap.put(bc.name, bc);}
ByteStreamWriter bsw=ByteStreamWriter.makeBSW(fname, overwrite, false, true);
bsw.println("Lane,SampleID,index,index2,# Reads,% of Hopped Reads,% of All Reads");
final ByteBuilder bb=new ByteBuilder();
for(String s : expectedSet) {
String sample=(sampleMap==null ? null : sampleMap.get(s));
lp.set(s);
bb.clear().append(lane).comma().append(sample==null ? "" : sample).comma();
bb.append(lp.parseByteArray(0)).comma();
bb.append(lp.parseByteArray(1)).comma();
Barcode bc=countMap.get(s);
long count=(bc==null ? 0 : bc.count());
bb.append(count).comma().comma().append(count*allMult,6).nl();
bsw.print(bb);
}
// bsw.poisonAndWait();
// assert(false);
long hoppedCount=0;
for(String left : leftSet) {
bb.clear().append(left).append(delimiter);
final int len=bb.length;
for(String right : rightSet) {
bb.setLength(len).append(right);
String pair=bb.toString();
if(!expectedSet.contains(pair)) {
Barcode bc=countMap.get(pair);
if(bc!=null) {
hoppedCount+=bc.count();
}
}
}
}
final float hoppedMult=1f/(Tools.max(1, hoppedCount));
ByteBuilder bb2=new ByteBuilder();
for(String left : leftSet) {
bb.clear().append(left).append(delimiter);
final int len=bb.length;
for(String right : rightSet) {
bb.setLength(len).append(right);
String pair=bb.toString();
if(!expectedSet.contains(pair)) {
Barcode bc=countMap.get(pair);
long count=(bc==null ? 0 : bc.count());
//String sample=(sampleMap==null ? null : sampleMap.get(s));
lp.set(pair);
bb2.clear().append(lane).comma().comma();
bb2.append(lp.parseByteArray(0)).comma();
bb2.append(lp.parseByteArray(1)).comma();
bb2.append(count).comma().append(count*hoppedMult, 6).comma();
bb2.append(count*allMult,6).nl();
bsw.print(bb2);
}
}
}
bsw.poison();
}
// if(fname==null) {return 0;}
// ArrayList<Barcode> list=new ArrayList<Barcode>(counts.size());
// Collections.sort(list);
//
// ByteStreamWriter bsw=ByteStreamWriter.makeBSW(fname, overwrite, false, true);
// if(delimiter<=0) {
// bsw.println("Lane,index,# Reads,% of Unknown Barcodes,% of All Reads");
// }else {
// bsw.println("Lane,index,index2,# Reads,% of Unknown Barcodes,% of All Reads");
// }
//
// long total=0;
// long expected=0;
// for(Barcode bc : counts) {
// total++;
// if(expectedSet.contains(bc.name)) {expected+=bc.count();}
// }
// final long unexpected=total-expected;
// final float invTotal=1f/Tools.max(1, total);
// final float invUnexpected=1f/Tools.max(1, unexpected);
//
// int printed=0;
// ByteBuilder bb=new ByteBuilder();
// for(Barcode bc : counts) {
// if(!expectedSet.contains(bc.name)) {
// if(printed>=lines) {break;}
// bb.clear().append(lane).comma();
// bc.appendIndex(bb, delimiter, 1).comma();
// if(delimiter>0) {bc.appendIndex(bb, delimiter, 2).comma();}
// bb.append(bc.count()).comma();
// bb.append(bc.count()*invUnexpected, 6).comma();
// bb.append(bc.count()*invTotal, 6).nl();
// bsw.print(bb);
// }
// }
// bsw.poison();
// return unexpected;
// }
// Quality_Metrics.csv
// Lane,SampleID,index,index2,ReadNumber,Yield,YieldQ30,QualityScoreSum,Mean Quality Score (PF),% Q30
// 7,529965,CCGTGCGGTC,CCCGATATGG,1,8405183819,7964042904,327093323760,38.92,0.95
// 7,529965,CCGTGCGGTC,CCCGATATGG,2,8405183819,7794032223,323344091568,38.47,0.93
// 7,529966,CACAGTTGTC,TAGGCAGGCT,1,7957915628,7552613802,309911557188,38.94,0.95
// ...
// 7,Undetermined,,,1,27221786230,23126098298,1001525318468,36.79,0.85
// 7,Undetermined,,,2,27221786230,22727595251,990581252636,36.39,0.83
int writeQualityMetrics(final Set<String> expectedSet, LinkedHashMap<String,String> sampleMap,
final byte delimiter, final String fname, final int lane, boolean overwrite) {
if(fname==null) {return 0;}
LinkedHashSet<String> set2=new LinkedHashSet<String>(expectedSet);
set2.remove(UNDETERMINED);
set2.add(UNDETERMINED);
ByteStreamWriter bsw=ByteStreamWriter.makeBSW(fname, overwrite, false, true);
bsw.println(QTag.header());
ByteBuilder bb=new ByteBuilder();
int lines=0;
for(String name : set2) {
String sample=(sampleMap==null ? null : sampleMap.get(name));
QTag[] tags=tagMap.get(name);
if(tags==null) {
tags=new QTag[] {new QTag(), new QTag()};
}
for(int pairnum=0; pairnum<tags.length && tags[pairnum]!=null; pairnum++) {
QTag tag=tags[pairnum];
bb.clear();
tag.appendTo(bb, lane, pairnum, sample==null ? UNDETERMINED : sample,
delimiter, name==UNDETERMINED ? "" : name);
bsw.print(bb.nl());
lines++;
}
}
bsw.poison();
return lines;
}
// Demultiplex_Stats.csv
// Lane,SampleID,Index,# Reads,# Perfect Index Reads,# One Mismatch Index Reads,# Two Mismatch Index Reads,% Reads,% Perfect Index Reads,% One Mismatch Index Reads,% Two Mismatch Index Reads
// 7,529965,CCGTGCGGTC-CCCGATATGG,55663469,49490086,6173383,0,0.0145,0.8891,0.1109,0.0000
// 7,529966,CACAGTTGTC-TAGGCAGGCT,52701428,48056108,4645320,0,0.0138,0.9119,0.0881,0.0000
// 7,529967,CCTCTTCAGA-GATCCAGTAG,48686653,44431620,4255033,0,0.0127,0.9126,0.0874,0.0000
void writeDemultiplexStats(String fname, HashMap<String, String> assignmentMap,
Set<String> expected, LinkedHashMap<String,String> sampleMap,
int lane, boolean overwrite) {
LinkedHashMap<String, long[]> hdistMap=new LinkedHashMap<String, long[]>(expected.size());
final int maxHDist=3;
for(String name : expected) {
hdistMap.put(name, new long[maxHDist+1]);
}
HashMap<String, Barcode> countMap=new HashMap<String, Barcode>(counts.size());
for(Barcode bc : counts) {countMap.put(bc.name, bc);}
for(Entry<String, String> e : assignmentMap.entrySet()) {
String key=e.getKey();
String value=e.getValue();
long[] distArray=hdistMap.get(value);
assert(distArray!=null) : value;
int dist=Tools.max(Barcode.hdistL(key, value), Barcode.hdistR(key, value));
Barcode bc=countMap.get(key);
long count=(bc==null ? 0 : bc.count());
distArray[Tools.min(maxHDist, dist)]+=count;
}
String header="Lane,SampleID,Index,# Reads,# Perfect Index Reads,"
+ "# One Mismatch Index Reads,# Two Mismatch Index Reads,"
+ "% Reads,% Perfect Index Reads,% One Mismatch Index Reads,% Two Mismatch Index Reads";
ByteStreamWriter bsw=ByteStreamWriter.makeBSW(fname, overwrite, false, true);
bsw.println(header);
ByteBuilder bb=new ByteBuilder();
final float totalMult=1f/totalR1;
for(String name : expected) {
String sample=(sampleMap==null ? null : sampleMap.get(name));
final long[] dist=hdistMap.get(name);
final long sum=Tools.sum(dist);
final float localMult=1f/sum;
bb.clear().append(lane);
bb.comma().append(sample==null ? "" : sample);
bb.comma().append(name.replace('+', '-'));
bb.comma().append(sum);
for(int i=0; i<=2; i++) {bb.comma().append(dist[i]);}
bb.comma().append(sum*totalMult, 4);
for(int i=0; i<=2; i++) {bb.comma().append(dist[i]*localMult, 4);}
bsw.print(bb.nl());
// System.err.println(totalR1+", "+sum+", "+Arrays.toString(dist)+", "+(sum*totalMult)+", "+bb);
}
{
long unknown=totalR1-totalR1Assigned;
String sample="Undetermined";
// final long[] dist=hdistMap.get(name);
final long sum=unknown;
// System.err.println(unknown+", "+totalR1+", "+totalR1Assigned);
final long[] dist=new long[] {sum, 0, 0};
final float localMult=1f/sum;
bb.clear().append(lane);
bb.comma().append(sample==null ? "" : sample);
bb.comma().append("");
bb.comma().append(sum);
for(int i=0; i<=2; i++) {bb.comma().append(dist[i]);}
bb.comma().append(sum*totalMult, 4);
for(int i=0; i<=2; i++) {bb.comma().append(dist[i]*localMult, 4);}
bsw.print(bb.nl());
}
bsw.poison();
}
// Demultiplex_Tile_Stats.csv
// Lane,SampleID,Index,Tile,# Reads,% Reads
// 7,529965,CCGTGCGGTC-CCCGATATGG,1101,29931,0.0000
// 7,529965,CCGTGCGGTC-CCCGATATGG,1102,7526,0.0000
// 7,529965,CCGTGCGGTC-CCCGATATGG,1103,67763,0.0000
void writeDemultiplexTileStats(String fname, LinkedHashMap<String,String> sampleMap,
Collection<String> expected, int lane, byte delimiter, boolean overwrite) {
if(tileMap==null) {return;}
if(expected==null) {expected=sampleMap.keySet();}
String header="Lane,SampleID,Index,Tile,# Reads,% Reads";
ByteStreamWriter bsw=ByteStreamWriter.makeBSW(fname, overwrite, false, true);
bsw.println(header);
ByteBuilder bb=new ByteBuilder();
ArrayList<Integer> tiles=new ArrayList<Integer>(tileMap.keySet());
Collections.sort(tiles);
final float invTotal=1f/Tools.max(1, totalR1);
// assert(false) : totalR1+", "+tileMap.size()+", "+expected.size();
for(Integer tile : tiles) {
LinkedHashMap<String, Barcode> tmap=tileMap.get(tile);
// for(Entry<String, Barcode> e : tmap.entrySet()) {
// Barcode bc=e.getValue();
for(String name : expected) {
Barcode bc=tmap.get(name);
if(bc!=null) {
String sampleID=(sampleMap==null ? null : sampleMap.get(bc.name));
bb.clear();
bb.append(lane);
bb.comma().append(sampleID);
Barcode.appendIndex(bb.comma(), delimiter, 1, bc.name);
if(delimiter>=0 && (byte)bc.name.indexOf(delimiter)>0) {
bb.dash();
Barcode.appendIndex(bb, delimiter, 2, bc.name);
}
bb.comma().append(tile);
bb.comma().append(bc.count());
bb.comma().append(bc.count()*invTotal, 4).nl();
bsw.print(bb);
}
}
}
bsw.poison();
}
void add(Read r, String barcode) {
if(barcode==null) {barcode=UNDETERMINED;}
QTag[] qtags=tagMap.get(barcode);
if(qtags==null) {
qtags=new QTag[] {new QTag(), new QTag()};
tagMap.put(barcode, qtags);
}
qtags[r.pairnum()].add(r);
String dest=(String) r.obj;
if(r.pairnum()==0) {
totalR1++;
totalR1Assigned+=(barcode==UNDETERMINED ? 0 : 1);
if(r.mate!=null) {
totalR2++;
totalR2Assigned+=(barcode==UNDETERMINED ? 0 : 1);
}
// assert(false) : (tileMap==null) +", "+dest+", "+barcode;
if(tileMap!=null && dest!=null) {
ihp.parse(r);
Integer tile=ihp.tile();
LinkedHashMap<String, Barcode> tmap=tileMap.get(tile);
if(tmap==null) {
tmap=new LinkedHashMap<String, Barcode>();
tileMap.put(tile, tmap);
}
Barcode bc=tmap.get(dest);
if(bc==null) {
bc=new Barcode(dest);
tmap.put(dest, bc);
}
assert(tileMap.size()>0 && tmap.size()>0) :
tileMap.size()+", "+tmap.size()+", "+tile+", "+dest+"\n"+ihp;
bc.increment(1);
}
}
}
LinkedHashMap<String, String> loadSampleMap(String fname,
byte barcodeDelimiter, boolean rcIndex1, boolean rcIndex2){
if(fname==null) {return null;}
LinkedHashMap<String, String> map=new LinkedHashMap<String, String>();
ByteFile bf=ByteFile.makeByteFile(fname, true);
LineParser1 lp=new LineParser1(',');
for(byte[] line=bf.nextLine(); line!=null; line=bf.nextLine()) {
if(!Tools.startsWith(line, '#')) {
lp.set(line);
if(lp.terms()<2) {
lp=new LineParser1('\t');
lp.set(line);
assert(lp.terms()>=2) : "Can't find delimiter in line '"+new String(line)+"'";
}
assert(lp.terms()>=2) : new String(line)+", "+lp;
String key=lp.parseString(0), value=lp.parseString(1);
key=BarcodeStats.fixBarcode(key, barcodeDelimiter, rcIndex1, rcIndex2);
map.put(key, value);
}
}
bf.close();
return map;
}
static class QTag {
int add(Read r) {
return add(r.quality);
}
int add(byte[] quals) {
reads++;
bases+=quals.length;
int q30=quals.length;
int q30Count=0;
// long sum=0;
for(int i=0; i<quals.length; i++) {
final int q=quals[i];
// sum+=q;
qsum.increment(i, q);
qcount.increment(i, 1);
if(q>=30) {
q30Count++;
}else if(i<q30){
q30=i;
}
}
q30PositionSum+=q30;
q30CountSum+=q30Count;
return q30;
}
long yield() {return bases;}
long yieldQ30() {return /*q30PositionSum*/q30CountSum;}
long qualityScoreSum() {return (long)qsum.sum();}
double meanQualityScore() {return qsum.sum()/bases;}
double fractionQ30() {return yieldQ30()/(double)yield();}
// Lane,SampleID,index,index2,ReadNumber,Yield,YieldQ30,QualityScoreSum,Mean Quality Score (PF),% Q30
ByteBuilder appendTo(ByteBuilder bb, int lane, int pairnum,
String sampleID, byte delimiter, String barcode) {
bb.append(lane);
bb.comma().append(sampleID);
Barcode.appendIndex(bb.comma(), delimiter, 1, barcode);
Barcode.appendIndex(bb.comma(), delimiter, 2, barcode);
bb.comma().append(pairnum+1);
bb.comma().append(yield());
bb.comma().append(yieldQ30());
bb.comma().append(qualityScoreSum());
bb.comma().append(meanQualityScore(),2);
bb.comma().append(fractionQ30(),2);
return bb;
}
static String header() {
return "Lane,SampleID,index,index2,ReadNumber,"
+ "Yield,YieldQ30,QualityScoreSum,Mean Quality Score (PF),% Q30";
}
long reads=0;
long bases=0;
long q30PositionSum=0;
long q30CountSum=0;
LongList qsum=new LongList();
LongList qcount=new LongList();
}
private long totalR1=0;
private long totalR2=0;
private long totalR1Assigned=0;
private long totalR2Assigned=0;
Collection<Barcode> counts=null;
final HashMap<String, QTag[]> tagMap=new HashMap<String, QTag[]>();
private final static String UNDETERMINED="Undetermined";;
LinkedHashMap<Integer, LinkedHashMap<String, Barcode>> tileMap=new LinkedHashMap<Integer, LinkedHashMap<String, Barcode>>();
final IlluminaHeaderParser2 ihp=new IlluminaHeaderParser2();
}
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