1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66 67 68 69 70 71 72 73 74 75 76 77 78 79 80 81 82 83 84 85 86 87 88 89 90 91 92 93 94 95 96 97 98 99 100 101 102 103 104 105 106 107 108 109 110 111 112 113 114 115 116 117 118 119 120 121 122 123 124 125 126 127 128 129 130 131 132 133 134 135 136 137 138 139 140 141 142 143 144 145 146 147 148 149 150 151 152 153 154 155 156 157 158 159 160 161 162 163 164 165 166 167 168 169 170 171 172 173 174 175 176 177 178 179 180 181 182 183 184 185 186 187 188 189 190 191 192 193 194 195 196 197 198 199 200 201 202 203 204 205 206 207 208 209 210 211 212 213 214 215 216 217 218 219 220 221 222 223 224 225 226 227 228 229 230 231 232 233 234 235 236 237 238 239 240 241 242 243 244 245 246 247 248 249 250 251 252 253 254 255 256 257 258 259 260 261 262 263 264 265 266 267 268 269 270 271 272 273 274 275 276 277 278 279 280 281 282 283 284 285 286 287 288 289 290 291 292 293 294 295 296 297 298 299 300 301 302 303 304 305 306 307 308 309 310 311 312 313 314 315 316 317 318 319 320 321 322 323 324 325 326 327 328 329 330 331 332 333 334 335 336 337 338 339 340 341 342 343 344 345 346 347 348 349 350 351 352 353 354 355 356 357 358 359 360 361 362 363 364 365 366 367 368 369 370 371 372 373 374 375 376 377 378 379 380 381 382 383 384 385 386 387 388 389 390 391 392 393 394 395 396 397 398 399 400 401 402 403 404 405 406 407 408 409 410 411 412 413 414 415 416 417 418 419 420 421 422 423 424 425 426 427 428 429 430 431 432 433 434 435 436 437 438 439 440 441 442 443 444 445 446 447 448 449 450 451 452 453 454 455 456 457 458 459 460 461 462 463 464 465 466 467 468 469 470 471 472 473 474 475 476 477 478 479 480 481 482 483 484 485 486 487 488 489 490 491 492 493 494 495 496 497 498 499 500 501 502 503 504 505 506 507 508 509 510 511 512 513 514 515 516 517 518 519 520 521 522 523 524 525 526 527 528 529 530 531 532 533 534 535 536 537 538 539 540 541 542 543 544 545 546 547 548 549 550 551 552 553 554 555 556 557 558 559 560 561 562 563 564 565 566 567 568 569 570 571 572 573 574 575 576 577 578 579 580 581 582 583 584 585 586 587 588 589 590 591 592 593 594 595 596 597 598 599 600 601 602 603 604 605 606 607 608 609 610 611 612 613 614 615 616 617 618 619 620 621 622 623 624 625 626 627 628 629 630 631 632 633 634 635 636 637 638 639 640 641 642 643 644 645 646 647 648 649 650 651 652 653 654 655 656 657 658 659 660 661 662 663 664 665 666 667 668 669 670 671 672 673 674 675 676 677 678 679 680 681 682 683 684 685 686 687 688 689 690 691 692 693 694 695 696 697 698 699 700 701 702 703 704 705 706 707 708 709 710 711 712 713 714 715 716 717 718 719 720 721 722 723 724 725 726 727 728 729 730 731 732 733 734 735 736 737 738 739 740 741 742 743 744 745 746 747 748 749 750 751 752 753 754 755 756 757 758 759 760 761 762 763 764 765 766 767 768 769 770 771 772 773 774 775 776 777 778 779 780 781 782 783 784 785 786 787 788 789 790 791 792 793 794 795 796 797 798 799 800 801 802 803 804 805 806 807 808 809 810 811 812 813 814 815 816 817 818 819 820 821 822 823 824 825 826 827 828 829 830 831 832 833 834 835 836 837 838 839 840 841 842 843 844 845 846 847 848 849 850 851 852 853 854 855 856 857 858 859 860 861 862 863 864 865 866 867 868 869 870 871 872 873 874 875 876 877 878 879 880 881 882 883 884 885 886 887 888 889 890 891 892 893 894 895 896 897 898 899 900 901 902 903 904 905 906 907 908 909 910 911 912 913 914 915 916 917 918 919 920 921 922 923 924 925 926 927 928 929 930 931 932 933 934 935 936 937 938 939 940 941 942 943 944 945 946 947 948 949 950 951 952 953 954 955 956 957 958 959 960 961 962 963 964 965 966 967 968 969 970 971 972 973 974 975 976 977 978 979 980 981 982 983 984 985 986 987 988 989 990 991 992 993 994 995 996 997 998 999 1000 1001 1002 1003 1004 1005 1006 1007 1008 1009 1010 1011 1012 1013 1014 1015 1016 1017 1018 1019 1020 1021 1022 1023 1024 1025 1026 1027 1028 1029 1030 1031 1032 1033 1034 1035 1036 1037 1038 1039 1040 1041 1042 1043 1044 1045 1046 1047 1048 1049 1050 1051 1052 1053 1054 1055 1056 1057 1058 1059 1060 1061 1062 1063 1064 1065 1066 1067 1068 1069 1070 1071 1072 1073 1074 1075 1076 1077 1078 1079 1080 1081 1082 1083 1084 1085 1086 1087 1088 1089 1090 1091
|
/*
* ccode.cpp
* Mothur
*
* Created by westcott on 8/24/09.
* Copyright 2009 Schloss Lab. All rights reserved.
*
*/
#include "ccode.h"
#include "ignoregaps.h"
#include "eachgapdist.h"
//***************************************************************************************************************
Ccode::Ccode(string filename, string temp, bool f, string mask, int win, int numW, string o) : Chimera() {
try {
fastafile = filename;
outputDir = o;
templateFileName = temp; templateSeqs = readSeqs(temp);
setMask(mask);
filter = f;
window = win;
numWanted = numW;
distCalc = new eachGapDist();
decalc = new DeCalculator();
mapInfo = outputDir + m->getRootName(m->getSimpleName(fastafile)) + "mapinfo";
#ifdef USE_MPI
//char* inFileName = new char[mapInfo.length()];
//memcpy(inFileName, mapInfo.c_str(), mapInfo.length());
char inFileName[1024];
strcpy(inFileName, mapInfo.c_str());
int outMode=MPI_MODE_CREATE|MPI_MODE_WRONLY;
MPI_File_open(MPI_COMM_WORLD, inFileName, outMode, MPI_INFO_NULL, &outMap); //comm, filename, mode, info, filepointer
//delete inFileName;
int pid;
MPI_Comm_rank(MPI_COMM_WORLD, &pid); //find out who we are
if (pid == 0) {
string outString = "Place in masked, filtered and trimmed sequence\tPlace in original alignment\n";
MPI_Status status;
int length = outString.length();
char* buf2 = new char[length];
memcpy(buf2, outString.c_str(), length);
MPI_File_write_shared(outMap, buf2, length, MPI_CHAR, &status);
delete buf2;
}
#else
ofstream out2;
m->openOutputFile(mapInfo, out2);
out2 << "Place in masked, filtered and trimmed sequence\tPlace in original alignment" << endl;
out2.close();
#endif
}
catch(exception& e) {
m->errorOut(e, "Ccode", "Ccode");
exit(1);
}
}
//***************************************************************************************************************
Ccode::~Ccode() {
delete distCalc;
delete decalc;
#ifdef USE_MPI
MPI_File_close(&outMap);
#endif
}
//***************************************************************************************************************
Sequence Ccode::print(ostream& out, ostream& outAcc) {
try {
ofstream out2;
m->openOutputFileAppend(mapInfo, out2);
out2 << querySeq->getName() << endl;
for (it = spotMap.begin(); it!= spotMap.end(); it++) {
out2 << it->first << '\t' << it->second << endl;
}
out2.close();
out << querySeq->getName() << endl << endl << "Reference sequences used and distance to query:" << endl;
for (int j = 0; j < closest.size(); j++) {
out << closest[j].seq->getName() << '\t' << closest[j].dist << endl;
}
out << endl << endl;
//for each window
//window mapping info.
out << "Mapping information: ";
//you mask and did not filter
if ((seqMask != "") && (!filter)) { out << "mask and trim."; }
//you filtered and did not mask
if ((seqMask == "") && (filter)) { out << "filter and trim."; }
//you masked and filtered
if ((seqMask != "") && (filter)) { out << "mask, filter and trim."; }
out << endl << "Window\tStartPos\tEndPos" << endl;
it = trim.begin();
for (int k = 0; k < windows.size()-1; k++) {
out << k+1 << '\t' << spotMap[windows[k]-it->first] << '\t' << spotMap[windows[k]-it->first+windowSizes] << endl;
}
out << windows.size() << '\t' << spotMap[windows[windows.size()-1]-it->first] << '\t' << spotMap[it->second-it->first-1] << endl;
out << endl;
out << "Window\tAvgQ\t(sdQ)\tAvgR\t(sdR)\tRatio\tAnova" << endl;
for (int k = 0; k < windows.size(); k++) {
float ds = averageQuery[k] / averageRef[k];
out << k+1 << '\t' << averageQuery[k] << '\t' << sdQuery[k] << '\t' << averageRef[k] << '\t'<< sdRef[k] << '\t' << ds << '\t' << anova[k] << endl;
}
out << endl;
//varRef
//varQuery
/* F test for differences among variances.
* varQuery is expected to be higher or similar than varRef */
//float fs = varQuery[query] / varRef[query]; /* F-Snedecor, test for differences of variances */
bool results = false;
//confidence limit, t - Student, anova
out << "Window\tConfidenceLimit\tt-Student\tAnova" << endl;
for (int k = 0; k < windows.size(); k++) {
string temp = "";
if (isChimericConfidence[k]) { temp += "*\t"; }
else { temp += "\t"; }
if (isChimericTStudent[k]) { temp += "*\t"; }
else { temp += "\t"; }
if (isChimericANOVA[k]) { temp += "*\t"; }
else { temp += "\t"; }
out << k+1 << '\t' << temp << endl;
if (temp == "*\t*\t*\t") { results = true; }
}
out << endl;
if (results) {
m->mothurOut(querySeq->getName() + " was found have at least one chimeric window."); m->mothurOutEndLine();
outAcc << querySeq->getName() << endl;
}
//free memory
for (int i = 0; i < closest.size(); i++) { delete closest[i].seq; }
return *querySeq;
}
catch(exception& e) {
m->errorOut(e, "Ccode", "print");
exit(1);
}
}
#ifdef USE_MPI
//***************************************************************************************************************
Sequence Ccode::print(MPI_File& out, MPI_File& outAcc) {
try {
string outMapString = "";
outMapString += querySeq->getName() + "\n";
for (it = spotMap.begin(); it!= spotMap.end(); it++) {
outMapString += toString(it->first) + "\t" + toString(it->second) + "\n";
}
printMapping(outMapString);
outMapString = "";
string outString = "";
string outAccString = "";
outString += querySeq->getName() + "\n\nReference sequences used and distance to query:\n";
for (int j = 0; j < closest.size(); j++) {
outString += closest[j].seq->getName() + "\t" + toString(closest[j].dist) + "\n";
}
outString += "\n\nMapping information: ";
//for each window
//window mapping info.
//you mask and did not filter
if ((seqMask != "") && (!filter)) { outString += "mask and trim."; }
//you filtered and did not mask
if ((seqMask == "") && (filter)) { outString += "filter and trim."; }
//you masked and filtered
if ((seqMask != "") && (filter)) { outString += "mask, filter and trim."; }
outString += "\nWindow\tStartPos\tEndPos\n";
it = trim.begin();
for (int k = 0; k < windows.size()-1; k++) {
outString += toString(k+1) + "\t" + toString(spotMap[windows[k]-it->first]) + "\t" + toString(spotMap[windows[k]-it->first+windowSizes]) + "\n";
}
outString += toString(windows.size()) + "\t" + toString(spotMap[windows[windows.size()-1]-it->first]) + "\t" + toString(spotMap[it->second-it->first-1]) + "\n\n";
outString += "Window\tAvgQ\t(sdQ)\tAvgR\t(sdR)\tRatio\tAnova\n";
for (int k = 0; k < windows.size(); k++) {
float ds = averageQuery[k] / averageRef[k];
outString += toString(k+1) + "\t" + toString(averageQuery[k]) + "\t" + toString(sdQuery[k]) + "\t" + toString(averageRef[k]) + "\t" + toString(sdRef[k]) + "\t" + toString(ds) + "\t" + toString(anova[k]) + "\n";
}
//varRef
//varQuery
/* F test for differences among variances.
* varQuery is expected to be higher or similar than varRef */
//float fs = varQuery[query] / varRef[query]; /* F-Snedecor, test for differences of variances */
bool results = false;
//confidence limit, t - Student, anova
outString += "\nWindow\tConfidenceLimit\tt-Student\tAnova\n";
for (int k = 0; k < windows.size(); k++) {
string temp = "";
if (isChimericConfidence[k]) { temp += "*\t"; }
else { temp += "\t"; }
if (isChimericTStudent[k]) { temp += "*\t"; }
else { temp += "\t"; }
if (isChimericANOVA[k]) { temp += "*\t"; }
else { temp += "\t"; }
outString += toString(k+1) + "\t" + temp + "\n";
if (temp == "*\t*\t*\t") { results = true; }
}
outString += "\n";
MPI_Status status;
int length = outString.length();
char* buf2 = new char[length];
memcpy(buf2, outString.c_str(), length);
MPI_File_write_shared(out, buf2, length, MPI_CHAR, &status);
delete buf2;
if (results) {
m->mothurOut(querySeq->getName() + " was found have at least one chimeric window."); m->mothurOutEndLine();
outAccString += querySeq->getName() + "\n";
MPI_Status statusAcc;
length = outAccString.length();
char* buf = new char[length];
memcpy(buf, outAccString.c_str(), length);
MPI_File_write_shared(outAcc, buf, length, MPI_CHAR, &statusAcc);
delete buf;
}
//free memory
for (int i = 0; i < closest.size(); i++) { delete closest[i].seq; }
return *querySeq;
}
catch(exception& e) {
m->errorOut(e, "Ccode", "print");
exit(1);
}
}
//***************************************************************************************************************
int Ccode::printMapping(string& output) {
try {
MPI_Status status;
int length = output.length();
char* buf = new char[length];
memcpy(buf, output.c_str(), length);
MPI_File_write_shared(outMap, buf, length, MPI_CHAR, &status);
delete buf;
return 0;
}
catch(exception& e) {
m->errorOut(e, "Ccode", "printMapping");
exit(1);
}
}
#endif
//***************************************************************************************************************
int Ccode::getChimeras(Sequence* query) {
try {
closest.clear();
refCombo = 0;
sumRef.clear();
varRef.clear();
varQuery.clear();
sdRef.clear();
sdQuery.clear();
sumQuery.clear();
sumSquaredRef.clear();
sumSquaredQuery.clear();
averageRef.clear();
averageQuery.clear();
anova.clear();
isChimericConfidence.clear();
isChimericTStudent.clear();
isChimericANOVA.clear();
trim.clear();
spotMap.clear();
windowSizes = window;
windows.clear();
querySeq = query;
//find closest matches to query
closest = findClosest(query, numWanted);
if (m->control_pressed) { return 0; }
//initialize spotMap
for (int i = 0; i < query->getAligned().length(); i++) { spotMap[i] = i; }
//mask sequences if the user wants to
if (seqMask != "") {
decalc->setMask(seqMask);
decalc->runMask(query);
//mask closest
for (int i = 0; i < closest.size(); i++) { decalc->runMask(closest[i].seq); }
spotMap = decalc->getMaskMap();
}
if (filter) {
vector<Sequence*> temp;
for (int i = 0; i < closest.size(); i++) { temp.push_back(closest[i].seq); }
temp.push_back(query);
createFilter(temp, 0.5);
for (int i = 0; i < temp.size(); i++) {
if (m->control_pressed) { return 0; }
runFilter(temp[i]);
}
//update spotMap
map<int, int> newMap;
int spot = 0;
for (int i = 0; i < filterString.length(); i++) {
if (filterString[i] == '1') {
//add to newMap
newMap[spot] = spotMap[i];
spot++;
}
}
spotMap = newMap;
}
//trim sequences - this follows ccodes remove_extra_gaps
trimSequences(query);
if (m->control_pressed) { return 0; }
//windows are equivalent to words - ccode paper recommends windows are between 5% and 20% on alignment length().
//Our default will be 10% and we will warn if user tries to use a window above or below these recommendations
windows = findWindows();
if (m->control_pressed) { return 0; }
//remove sequences that are more than 20% different and less than 0.5% different - may want to allow user to specify this later
removeBadReferenceSeqs(closest);
if (m->control_pressed) { return 0; }
//find the averages for each querys references
getAverageRef(closest); //fills sumRef, averageRef, sumSquaredRef and refCombo.
getAverageQuery(closest, query); //fills sumQuery, averageQuery, sumSquaredQuery.
if (m->control_pressed) { return 0; }
//find the averages for each querys references
findVarianceRef(); //fills varRef and sdRef also sets minimum error rate to 0.001 to avoid divide by 0.
if (m->control_pressed) { return 0; }
//find the averages for the query
findVarianceQuery(); //fills varQuery and sdQuery also sets minimum error rate to 0.001 to avoid divide by 0.
if (m->control_pressed) { return 0; }
determineChimeras(); //fills anova, isChimericConfidence, isChimericTStudent and isChimericANOVA.
if (m->control_pressed) { return 0; }
return 0;
}
catch(exception& e) {
m->errorOut(e, "Ccode", "getChimeras");
exit(1);
}
}
/***************************************************************************************************************/
//ccode algo says it does this to "Removes the initial and final gaps to avoid biases due to incomplete sequences."
void Ccode::trimSequences(Sequence* query) {
try {
int frontPos = 0; //should contain first position in all seqs that is not a gap character
int rearPos = query->getAligned().length();
//********find first position in closest seqs that is a non gap character***********//
//find first position all query seqs that is a non gap character
for (int i = 0; i < closest.size(); i++) {
string aligned = closest[i].seq->getAligned();
int pos = 0;
//find first spot in this seq
for (int j = 0; j < aligned.length(); j++) {
if (isalpha(aligned[j])) {
pos = j;
break;
}
}
//save this spot if it is the farthest
if (pos > frontPos) { frontPos = pos; }
}
//find first position all querySeq[query] that is a non gap character
string aligned = query->getAligned();
int pos = 0;
//find first spot in this seq
for (int j = 0; j < aligned.length(); j++) {
if (isalpha(aligned[j])) {
pos = j;
break;
}
}
//save this spot if it is the farthest
if (pos > frontPos) { frontPos = pos; }
//********find last position in closest seqs that is a non gap character***********//
for (int i = 0; i < closest.size(); i++) {
string aligned = closest[i].seq->getAligned();
int pos = aligned.length();
//find first spot in this seq
for (int j = aligned.length()-1; j >= 0; j--) {
if (isalpha(aligned[j])) {
pos = j;
break;
}
}
//save this spot if it is the farthest
if (pos < rearPos) { rearPos = pos; }
}
//find last position all querySeqs[query] that is a non gap character
aligned = query->getAligned();
pos = aligned.length();
//find first spot in this seq
for (int j = aligned.length()-1; j >= 0; j--) {
if (isalpha(aligned[j])) {
pos = j;
break;
}
}
//save this spot if it is the farthest
if (pos < rearPos) { rearPos = pos; }
//check to make sure that is not whole seq
if ((rearPos - frontPos - 1) <= 0) { m->mothurOut("Error, when I trim your sequences, the entire sequence is trimmed."); m->mothurOutEndLine(); exit(1); }
map<int, int> tempTrim;
tempTrim[frontPos] = rearPos;
//save trimmed locations
trim = tempTrim;
//update spotMask
map<int, int> newMap;
int spot = 0;
for (int i = frontPos; i < rearPos; i++) {
//add to newMap
newMap[spot] = spotMap[i];
spot++;
}
spotMap = newMap;
}
catch(exception& e) {
m->errorOut(e, "Ccode", "trimSequences");
exit(1);
}
}
/***************************************************************************************************************/
vector<int> Ccode::findWindows() {
try {
vector<int> win;
it = trim.begin();
int length = it->second - it->first;
//default is wanted = 10% of total length
if (windowSizes > length) {
m->mothurOut("You have slected a window larger than your sequence length after all filters, masks and trims have been done. I will use the default 10% of sequence length.");
windowSizes = length / 10;
}else if (windowSizes == 0) { windowSizes = length / 10; }
else if (windowSizes > (length * 0.20)) {
m->mothurOut("You have selected a window that is larger than 20% of your sequence length. This is not recommended, but I will continue anyway."); m->mothurOutEndLine();
}else if (windowSizes < (length * 0.05)) {
m->mothurOut("You have selected a window that is smaller than 5% of your sequence length. This is not recommended, but I will continue anyway."); m->mothurOutEndLine();
}
//save starting points of each window
for (int m = it->first; m < (it->second-windowSizes); m+=windowSizes) { win.push_back(m); }
//save last window
if (win[win.size()-1] < (it->first+length)) {
win.push_back(win[win.size()-1]+windowSizes); // ex. string length is 115, window is 25, without this you would get 0, 25, 50, 75
} //with this you would get 1,25,50,75,100
return win;
}
catch(exception& e) {
m->errorOut(e, "Ccode", "findWindows");
exit(1);
}
}
//***************************************************************************************************************
int Ccode::getDiff(string seqA, string seqB) {
try {
int numDiff = 0;
for (int i = 0; i < seqA.length(); i++) {
//if you are both not gaps
//if (isalpha(seqA[i]) && isalpha(seqA[i])) {
//are you different
if (seqA[i] != seqB[i]) {
int ok; /* ok=1 means equivalent base. Checks for degenerate bases */
/* the char in base_a and base_b have been checked and they are different */
if ((seqA[i] == 'N') && (seqB[i] != '-')) ok = 1;
else if ((seqB[i] == 'N') && (seqA[i] != '-')) ok = 1;
else if ((seqA[i] == 'Y') && ((seqB[i] == 'C') || (seqB[i] == 'T'))) ok = 1;
else if ((seqB[i] == 'Y') && ((seqA[i] == 'C') || (seqA[i] == 'T'))) ok = 1;
else if ((seqA[i] == 'R') && ((seqB[i] == 'G') || (seqB[i] == 'A'))) ok = 1;
else if ((seqB[i] == 'R') && ((seqA[i] == 'G') || (seqA[i] == 'A'))) ok = 1;
else if ((seqA[i] == 'S') && ((seqB[i] == 'C') || (seqB[i] == 'G'))) ok = 1;
else if ((seqB[i] == 'S') && ((seqA[i] == 'C') || (seqA[i] == 'G'))) ok = 1;
else if ((seqA[i] == 'W') && ((seqB[i] == 'T') || (seqB[i] == 'A'))) ok = 1;
else if ((seqB[i] == 'W') && ((seqA[i] == 'T') || (seqA[i] == 'A'))) ok = 1;
else if ((seqA[i] == 'M') && ((seqB[i] == 'A') || (seqB[i] == 'C'))) ok = 1;
else if ((seqB[i] == 'M') && ((seqA[i] == 'A') || (seqA[i] == 'C'))) ok = 1;
else if ((seqA[i] == 'K') && ((seqB[i] == 'T') || (seqB[i] == 'G'))) ok = 1;
else if ((seqB[i] == 'K') && ((seqA[i] == 'T') || (seqA[i] == 'G'))) ok = 1;
else if ((seqA[i] == 'V') && ((seqB[i] == 'C') || (seqB[i] == 'A') || (seqB[i] == 'G'))) ok = 1;
else if ((seqB[i] == 'V') && ((seqA[i] == 'C') || (seqA[i] == 'A') || (seqA[i] == 'G'))) ok = 1;
else if ((seqA[i] == 'H') && ((seqB[i] == 'T') || (seqB[i] == 'A') || (seqB[i] == 'C'))) ok = 1;
else if ((seqB[i] == 'H') && ((seqA[i] == 'T') || (seqA[i] == 'A') || (seqA[i] == 'C'))) ok = 1;
else if ((seqA[i] == 'D') && ((seqB[i] == 'T') || (seqB[i] == 'A') || (seqB[i] == 'G'))) ok = 1;
else if ((seqB[i] == 'D') && ((seqA[i] == 'T') || (seqA[i] == 'A') || (seqA[i] == 'G'))) ok = 1;
else if ((seqA[i] == 'B') && ((seqB[i] == 'C') || (seqB[i] == 'T') || (seqB[i] == 'G'))) ok = 1;
else if ((seqB[i] == 'B') && ((seqA[i] == 'C') || (seqA[i] == 'T') || (seqA[i] == 'G'))) ok = 1;
else ok = 0; /* the bases are different and not equivalent */
//check if they are both blanks
if ((seqA[i] == '.') && (seqB[i] == '-')) ok = 1;
else if ((seqB[i] == '.') && (seqA[i] == '-')) ok = 1;
if (ok == 0) { numDiff++; }
}
//}
}
return numDiff;
}
catch(exception& e) {
m->errorOut(e, "Ccode", "getDiff");
exit(1);
}
}
//***************************************************************************************************************
//tried to make this look most like ccode original implementation
void Ccode::removeBadReferenceSeqs(vector<SeqDist>& seqs) {
try {
vector< vector<int> > numDiffBases;
numDiffBases.resize(seqs.size());
//initialize to 0
for (int i = 0; i < numDiffBases.size(); i++) { numDiffBases[i].resize(seqs.size(),0); }
it = trim.begin();
int length = it->second - it->first;
//calc differences from each sequence to everyother seq in the set
for (int i = 0; i < seqs.size(); i++) {
string seqA = seqs[i].seq->getAligned().substr(it->first, length);
//so you don't calc i to j and j to i since they are the same
for (int j = 0; j < i; j++) {
string seqB = seqs[j].seq->getAligned().substr(it->first, length);
//compare strings
int numDiff = getDiff(seqA, seqB);
numDiffBases[i][j] = numDiff;
numDiffBases[j][i] = numDiff;
}
}
//initailize remove to 0
vector<int> remove; remove.resize(seqs.size(), 0);
float top = ((20*length) / (float) 100);
float bottom = ((0.5*length) / (float) 100);
//check each numDiffBases and if any are higher than threshold set remove to 1 so you can remove those seqs from the closest set
for (int i = 0; i < numDiffBases.size(); i++) {
for (int j = 0; j < i; j++) {
//are you more than 20% different
if (numDiffBases[i][j] > top) { remove[j] = 1; }
//are you less than 0.5% different
if (numDiffBases[i][j] < bottom) { remove[j] = 1; }
}
}
int numSeqsLeft = 0;
//count seqs that are not going to be removed
for (int i = 0; i < remove.size(); i++) {
if (remove[i] == 0) { numSeqsLeft++; }
}
//if you have enough then remove bad ones
if (numSeqsLeft >= 3) {
vector<SeqDist> goodSeqs;
//remove bad seqs
for (int i = 0; i < remove.size(); i++) {
if (remove[i] == 0) {
goodSeqs.push_back(seqs[i]);
}
}
seqs = goodSeqs;
}else { //warn, but dont remove any
m->mothurOut(querySeq->getName() + " does not have an adaquate number of reference sequences that are within 20% and 0.5% similarity. I will continue, but please check."); m->mothurOutEndLine();
}
}
catch(exception& e) {
m->errorOut(e, "Ccode", "removeBadReferenceSeqs");
exit(1);
}
}
//***************************************************************************************************************
//makes copy of templateseq for filter
vector<SeqDist> Ccode::findClosest(Sequence* q, int numWanted) {
try{
vector<SeqDist> topMatches;
Sequence query = *(q);
//calc distance to each sequence in template seqs
for (int i = 0; i < templateSeqs.size(); i++) {
Sequence ref = *(templateSeqs[i]);
//find overall dist
distCalc->calcDist(query, ref);
float dist = distCalc->getDist();
//save distance
SeqDist temp;
temp.seq = new Sequence(templateSeqs[i]->getName(), templateSeqs[i]->getAligned());
temp.dist = dist;
topMatches.push_back(temp);
}
sort(topMatches.begin(), topMatches.end(), compareSeqDist);
for (int i = numWanted; i < topMatches.size(); i++) { delete topMatches[i].seq; }
topMatches.resize(numWanted);
return topMatches;
}
catch(exception& e) {
m->errorOut(e, "Ccode", "findClosestSides");
exit(1);
}
}
/**************************************************************************************************/
//find the distances from each reference sequence to every other reference sequence for each window for this query
void Ccode::getAverageRef(vector<SeqDist> ref) {
try {
vector< vector< vector<int> > > diffs; //diffs[0][1][2] is the number of differences between ref seq 0 and ref seq 1 at window 2.
//initialize diffs vector
diffs.resize(ref.size());
for (int i = 0; i < diffs.size(); i++) {
diffs[i].resize(ref.size());
for (int j = 0; j < diffs[i].size(); j++) {
diffs[i][j].resize(windows.size(), 0);
}
}
it = trim.begin();
//find the distances from each reference sequence to every other reference sequence for each window for this query
for (int i = 0; i < ref.size(); i++) {
string refI = ref[i].seq->getAligned();
//j<i, so you don't find distances from i to j and then j to i.
for (int j = 0; j < i; j++) {
string refJ = ref[j].seq->getAligned();
for (int k = 0; k < windows.size(); k++) {
string refIWindowk, refJWindowk;
if (k < windows.size()-1) {
//get window strings
refIWindowk = refI.substr(windows[k], windowSizes);
refJWindowk = refJ.substr(windows[k], windowSizes);
}else { //last window may be smaller than rest - see findwindows
//get window strings
refIWindowk = refI.substr(windows[k], (it->second-windows[k]));
refJWindowk = refJ.substr(windows[k], (it->second-windows[k]));
}
//find differences
int diff = getDiff(refIWindowk, refJWindowk);
//save differences in [i][j][k] and [j][i][k] since they are the same
diffs[i][j][k] = diff;
diffs[j][i][k] = diff;
}//k
}//j
}//i
//initialize sumRef for this query
sumRef.resize(windows.size(), 0);
sumSquaredRef.resize(windows.size(), 0);
averageRef.resize(windows.size(), 0);
//find the sum of the differences for hte reference sequences
for (int i = 0; i < diffs.size(); i++) {
for (int j = 0; j < i; j++) {
//increment this querys reference sequences combos
refCombo++;
for (int k = 0; k < diffs[i][j].size(); k++) {
sumRef[k] += diffs[i][j][k];
sumSquaredRef[k] += (diffs[i][j][k]*diffs[i][j][k]);
}//k
}//j
}//i
//find the average of the differences for the references for each window
for (int i = 0; i < windows.size(); i++) {
averageRef[i] = sumRef[i] / (float) refCombo;
}
}
catch(exception& e) {
m->errorOut(e, "Ccode", "getAverageRef");
exit(1);
}
}
/**************************************************************************************************/
void Ccode::getAverageQuery (vector<SeqDist> ref, Sequence* query) {
try {
vector< vector<int> > diffs; //diffs[1][2] is the number of differences between querySeqs[query] and ref seq 1 at window 2.
//initialize diffs vector
diffs.resize(ref.size());
for (int j = 0; j < diffs.size(); j++) {
diffs[j].resize(windows.size(), 0);
}
it = trim.begin();
string refQuery = query->getAligned();
//j<i, so you don't find distances from i to j and then j to i.
for (int j = 0; j < ref.size(); j++) {
string refJ = ref[j].seq->getAligned();
for (int k = 0; k < windows.size(); k++) {
string QueryWindowk, refJWindowk;
if (k < windows.size()-1) {
//get window strings
QueryWindowk = refQuery.substr(windows[k], windowSizes);
refJWindowk = refJ.substr(windows[k], windowSizes);
}else { //last window may be smaller than rest - see findwindows
//get window strings
QueryWindowk = refQuery.substr(windows[k], (it->second-windows[k]));
refJWindowk = refJ.substr(windows[k], (it->second-windows[k]));
}
//find differences
int diff = getDiff(QueryWindowk, refJWindowk);
//save differences
diffs[j][k] = diff;
}//k
}//j
//initialize sumRef for this query
sumQuery.resize(windows.size(), 0);
sumSquaredQuery.resize(windows.size(), 0);
averageQuery.resize(windows.size(), 0);
//find the sum of the differences
for (int j = 0; j < diffs.size(); j++) {
for (int k = 0; k < diffs[j].size(); k++) {
sumQuery[k] += diffs[j][k];
sumSquaredQuery[k] += (diffs[j][k]*diffs[j][k]);
}//k
}//j
//find the average of the differences for the references for each window
for (int i = 0; i < windows.size(); i++) {
averageQuery[i] = sumQuery[i] / (float) ref.size();
}
}
catch(exception& e) {
m->errorOut(e, "Ccode", "getAverageQuery");
exit(1);
}
}
/**************************************************************************************************/
void Ccode::findVarianceRef() {
try {
varRef.resize(windows.size(), 0);
sdRef.resize(windows.size(), 0);
//for each window
for (int i = 0; i < windows.size(); i++) {
varRef[i] = (sumSquaredRef[i] - ((sumRef[i]*sumRef[i])/(float)refCombo)) / (float)(refCombo-1);
sdRef[i] = sqrt(varRef[i]);
//set minimum error rate to 0.001 - to avoid potential divide by zero - not sure if this is necessary but it follows ccode implementation
if (averageRef[i] < 0.001) { averageRef[i] = 0.001; }
if (sumRef[i] < 0.001) { sumRef[i] = 0.001; }
if (varRef[i] < 0.001) { varRef[i] = 0.001; }
if (sumSquaredRef[i] < 0.001) { sumSquaredRef[i] = 0.001; }
if (sdRef[i] < 0.001) { sdRef[i] = 0.001; }
}
}
catch(exception& e) {
m->errorOut(e, "Ccode", "findVarianceRef");
exit(1);
}
}
/**************************************************************************************************/
void Ccode::findVarianceQuery() {
try {
varQuery.resize(windows.size(), 0);
sdQuery.resize(windows.size(), 0);
//for each window
for (int i = 0; i < windows.size(); i++) {
varQuery[i] = (sumSquaredQuery[i] - ((sumQuery[i]*sumQuery[i])/(float) closest.size())) / (float) (closest.size()-1);
sdQuery[i] = sqrt(varQuery[i]);
//set minimum error rate to 0.001 - to avoid potential divide by zero - not sure if this is necessary but it follows ccode implementation
if (averageQuery[i] < 0.001) { averageQuery[i] = 0.001; }
if (sumQuery[i] < 0.001) { sumQuery[i] = 0.001; }
if (varQuery[i] < 0.001) { varQuery[i] = 0.001; }
if (sumSquaredQuery[i] < 0.001) { sumSquaredQuery[i] = 0.001; }
if (sdQuery[i] < 0.001) { sdQuery[i] = 0.001; }
}
}
catch(exception& e) {
m->errorOut(e, "Ccode", "findVarianceQuery");
exit(1);
}
}
/**************************************************************************************************/
void Ccode::determineChimeras() {
try {
isChimericConfidence.resize(windows.size(), false);
isChimericTStudent.resize(windows.size(), false);
isChimericANOVA.resize(windows.size(), false);
anova.resize(windows.size());
//for each window
for (int i = 0; i < windows.size(); i++) {
//get confidence limits
float t = getT(closest.size()-1); //how many seqs you are comparing to this querySeq
float dsUpper = (averageQuery[i] + (t * sdQuery[i])) / averageRef[i];
float dsLower = (averageQuery[i] - (t * sdQuery[i])) / averageRef[i];
if ((dsUpper > 1.0) && (dsLower > 1.0) && (averageQuery[i] > averageRef[i])) { /* range does not include 1 */
isChimericConfidence[i] = true; /* significantly higher at P<0.05 */
}
//student t test
int degreeOfFreedom = refCombo + closest.size() - 2;
float denomForT = (((refCombo-1) * varQuery[i] + (closest.size() - 1) * varRef[i]) / (float) degreeOfFreedom) * ((refCombo + closest.size()) / (float) (refCombo * closest.size())); /* denominator, without sqrt(), for ts calculations */
float ts = fabs((averageQuery[i] - averageRef[i]) / (sqrt(denomForT))); /* value of ts for t-student test */
t = getT(degreeOfFreedom);
if ((ts >= t) && (averageQuery[i] > averageRef[i])) {
isChimericTStudent[i] = true; /* significantly higher at P<0.05 */
}
//anova test
float value1 = sumQuery[i] + sumRef[i];
float value2 = sumSquaredQuery[i] + sumSquaredRef[i];
float value3 = ((sumQuery[i]*sumQuery[i]) / (float) (closest.size())) + ((sumRef[i] * sumRef[i]) / (float) refCombo);
float value4 = (value1 * value1) / ( (float) (closest.size() + refCombo) );
float value5 = value2 - value4;
float value6 = value3 - value4;
float value7 = value5 - value6;
float value8 = value7 / ((float) degreeOfFreedom);
float anovaValue = value6 / value8;
float f = getF(degreeOfFreedom);
if ((anovaValue >= f) && (averageQuery[i] > averageRef[i])) {
isChimericANOVA[i] = true; /* significant P<0.05 */
}
if (isnan(anovaValue) || isinf(anovaValue)) { anovaValue = 0.0; }
anova[i] = anovaValue;
}
}
catch(exception& e) {
m->errorOut(e, "Ccode", "determineChimeras");
exit(1);
}
}
/**************************************************************************************************/
float Ccode::getT(int numseq) {
try {
float tvalue = 0;
/* t-student critical values for different degrees of freedom and alpha 0.1 in one-tail tests (equivalent to 0.05) */
if (numseq > 120) tvalue = 1.645;
else if (numseq > 60) tvalue = 1.658;
else if (numseq > 40) tvalue = 1.671;
else if (numseq > 30) tvalue = 1.684;
else if (numseq > 29) tvalue = 1.697;
else if (numseq > 28) tvalue = 1.699;
else if (numseq > 27) tvalue = 1.701;
else if (numseq > 26) tvalue = 1.703;
else if (numseq > 25) tvalue = 1.706;
else if (numseq > 24) tvalue = 1.708;
else if (numseq > 23) tvalue = 1.711;
else if (numseq > 22) tvalue = 1.714;
else if (numseq > 21) tvalue = 1.717;
else if (numseq > 20) tvalue = 1.721;
else if (numseq > 19) tvalue = 1.725;
else if (numseq > 18) tvalue = 1.729;
else if (numseq > 17) tvalue = 1.734;
else if (numseq > 16) tvalue = 1.740;
else if (numseq > 15) tvalue = 1.746;
else if (numseq > 14) tvalue = 1.753;
else if (numseq > 13) tvalue = 1.761;
else if (numseq > 12) tvalue = 1.771;
else if (numseq > 11) tvalue = 1.782;
else if (numseq > 10) tvalue = 1.796;
else if (numseq > 9) tvalue = 1.812;
else if (numseq > 8) tvalue = 1.833;
else if (numseq > 7) tvalue = 1.860;
else if (numseq > 6) tvalue = 1.895;
else if (numseq > 5) tvalue = 1.943;
else if (numseq > 4) tvalue = 2.015;
else if (numseq > 3) tvalue = 2.132;
else if (numseq > 2) tvalue = 2.353;
else if (numseq > 1) tvalue = 2.920;
else if (numseq <= 1) {
m->mothurOut("Two or more reference sequences are required, your data will be flawed.\n"); m->mothurOutEndLine();
}
return tvalue;
}
catch(exception& e) {
m->errorOut(e, "Ccode", "getT");
exit(1);
}
}
/**************************************************************************************************/
float Ccode::getF(int numseq) {
try {
float fvalue = 0;
/* F-Snedecor critical values for v1=1 and different degrees of freedom v2 and alpha 0.05 */
if (numseq > 120) fvalue = 3.84;
else if (numseq > 60) fvalue = 3.92;
else if (numseq > 40) fvalue = 4.00;
else if (numseq > 30) fvalue = 4.08;
else if (numseq > 29) fvalue = 4.17;
else if (numseq > 28) fvalue = 4.18;
else if (numseq > 27) fvalue = 4.20;
else if (numseq > 26) fvalue = 4.21;
else if (numseq > 25) fvalue = 4.23;
else if (numseq > 24) fvalue = 4.24;
else if (numseq > 23) fvalue = 4.26;
else if (numseq > 22) fvalue = 4.28;
else if (numseq > 21) fvalue = 4.30;
else if (numseq > 20) fvalue = 4.32;
else if (numseq > 19) fvalue = 4.35;
else if (numseq > 18) fvalue = 4.38;
else if (numseq > 17) fvalue = 4.41;
else if (numseq > 16) fvalue = 4.45;
else if (numseq > 15) fvalue = 4.49;
else if (numseq > 14) fvalue = 4.54;
else if (numseq > 13) fvalue = 4.60;
else if (numseq > 12) fvalue = 4.67;
else if (numseq > 11) fvalue = 4.75;
else if (numseq > 10) fvalue = 4.84;
else if (numseq > 9) fvalue = 4.96;
else if (numseq > 8) fvalue = 5.12;
else if (numseq > 7) fvalue = 5.32;
else if (numseq > 6) fvalue = 5.59;
else if (numseq > 5) fvalue = 5.99;
else if (numseq > 4) fvalue = 6.61;
else if (numseq > 3) fvalue = 7.71;
else if (numseq > 2) fvalue = 10.1;
else if (numseq > 1) fvalue = 18.5;
else if (numseq > 0) fvalue = 161;
else if (numseq <= 0) {
m->mothurOut("Two or more reference sequences are required, your data will be flawed.\n"); m->mothurOutEndLine();
}
return fvalue;
}
catch(exception& e) {
m->errorOut(e, "Ccode", "getF");
exit(1);
}
}
//***************************************************************************************************************
|