732:
572:
100:
330:
media. The ones used are mostly green or light blue colour, and are chemically unstable. For that reason, unstabilized cyanine discs are unsuitable for archival CD and DVD use. Recent cyanine discs contain stabilizers, typically a metal atom bonded to the cyanine molecule, that slow the deterioration
846:
experiments so that samples from two sources can be mixed and run together through the separation process. This eliminates variations due to differing experimental conditions that are inevitable if the samples were run separately. These variations make it extremely difficult, if not impossible, to
834:
Cy5 is sensitive to its electronic environment. Changes in the conformation of the protein it is attached to will produce either enhancement or quenching of the emission. The rate of this change can be measured to determine enzyme kinetic parameters. The dyes can be used for similar purposes in
598:, and in solution. Cy5 became a popular replacement for far red fluorescent dyes because of its high extinction coefficient (as small as 1 nanomol can be detected in gel electrophoresis by naked eye) and its fluorophore emission maximum in the red region, where many
1234:
719:, Sulfo Cy dyes, Seta dyes, IRIS dyes from Cyanine Technologies and others can be used interchangeably with Cy dyes in most biochemical applications, with claimed improvements in solubility, fluorescence, or photostability.
347:
For applications to biotechnology, special cyanine dyes are synthesized from 2, 3, 5 or 7-methine structures with reactive groups on either one or both of the nitrogen ends so that they can be chemically linked to either
589:
is fluorescent in the far-red region (~650 excitation, 670 nm emission). Cy3 can be detected by various fluorometers, imagers, and microscopes with standard filters for
Tetramethylrhodamine (TRITC). Due to its high
802:
groups. This is done through incorporating aminoallyl-modified nucleotides during synthesis reactions. A good ratio is a label every 60 bases such that the labels are not too close to each other, which would result in
1197:
Umezawa K, Matsui A, Nakamura Y, Citterio D, Suzuki K (2009). "Bright, color-tunable fluorescent dyes in the Vis/NIR region: establishment of new "tailor-made" multicolor fluorophores based on borondipyrromethene".
625:) to avoid background contamination. They are thus able to easily distinguish colors from Cy3 and from Cy5, and also able to quantify the amount of Cy3 and Cy5 labeling in one sample (multiparametric detection).
670:
The Cy3 and Cy5 nomenclature was first proposed by Ernst, et al. in 1989, and is non-standard since it gives no hint of their chemical structures. In the original paper the number designated the count of the
722:
While patent protection for the standard Cy series of dyes has lapsed, the trademarked Cy naming remains in place. Consequently, dyes that are identical to Cy dyes, but called different names, are now sold.
743:
Cyanine dyes are used to label proteins, antibodies, peptides, nucleic acid probes, and any kind of other biomolecules to be used in a variety of fluorescence detection techniques:
383:
Cyanine dyes are available with different modifications such as methyl, ethyl or butyl substituents, carboxyl, acetylmethoxy, and sulfo groups which alter their hydrophilicity.
1489:
658:–Cyanine dyes bear one or two sulfo groups, rendering the Cy dye water-soluble, but tri- and quadri-sulfonated forms are available for even higher water solubility.
893:
Kvach, Maksim V.; Ustinov, Alexey V.; Stepanova, Irina A.; Malakhov, Andrei D.; Skorobogatyi, Mikhail V.; Shmanai, Vadim V.; Korshun, Vladimir A. (2008).
992:
1488:
Osterman IA, Ustinov AV, Evdokimov DV, Korshun VA, Sergiev PV, Serebryakova MV, Demina IA, Galyamina MA, Govorun VM, Dontsova OA (Jan 2013).
1162:
Mujumdar B, Ernst A, Mujumdar SR, Lewis CJ, Waggoner AS (Mar 1993). "Cyanine dye labeling reagents: Sulfoindocyanine succinimidyl esters".
1445:
Unlü M, Morgan ME, Minden JS (Oct 1997). "Difference gel electrophoresis: a single gel method for detecting changes in protein extracts".
964:
Advanced
Fluorescence Reporters in Chemistry and Biology I: Fundamentals and Molecular Design Volume 8 of Springer Series on Fluorescence
847:
use computers to automate the acquisition of the data after the separation is complete. Using these dyes makes the automation trivial.
310:
Cyanines were first synthesized over a century ago. They were originally used, and still are, to increase the sensitivity range of
1567:
1345:
1409:
1528:
836:
373:
555:
Because they yield brighter and more stable fluorescence, cyanines can advantageously replace conventional dyes such as
1009:
331:
significantly. These discs are often rated with an archival life of 75 years or more. The other dyes used in CD-Rs are
1392:
Armitage, Bruce A. (27 January 2005). "Cyanine Dye–DNA Interactions: Intercalation, Groove
Binding, and Aggregation".
1235:"Cyanine dyes in biophysical research: the photophysics of polymethine fluorescent dyes in biomolecular environments"
971:
357:
648:
is a near-IR fluor that is invisible to the naked eye (excitation/emission maximum 750/776 nm). It is used in
894:
707:
Many analogs of standard Cy 2 / 3 / 3.5 / 5 / 5.5 / 7 / 7.5 dyes were developed, using diverse modification:
83:. Nevertheless, there is a wide literature on both their synthesis and uses, and cyanines are common in some
679:
were unspecified. Due to this ambiguity various structures are designated Cy3 and Cy5 in the literature. The
662:
is another modification that confers hydrophilicity, not only to the dye but also to the labeled conjugate.
591:
356:
molecules. Labeling is done for visualization and quantification purposes. Biological applications include
783:
is labeled with either Cy3 or Cy5 that has been synthesized to carry an N-hydroxysuccinimidyl ester (
1431:
1309:
1097:"Genome-wide analysis demonstrates conserved localization of messenger RNAs to mitotic microtubules"
804:
372:
such as RNA localization, molecular interaction studies by fluorescence resonance energy transfer (
42:
642:
is a near-infrared (IR) fluorescence-emitting dye (excitation/emission maximum 678/694 nm).
80:
959:
752:
1562:
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816:
784:
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361:
8:
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65:
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1096:
855:
The word cyanin is from the
English word "cyan", which conventionally means a shade of
688:
271:
268:
1512:
1462:
1405:
1289:
1262:
1254:
1215:
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914:
57:
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1454:
1397:
1274:
1246:
1207:
1171:
1116:
1108:
1041:
997:
940:
906:
895:"A Convenient Synthesis of Cyanine Dyes: Reagents for the Labeling of Biomolecules"
537:
1474:
1001:
602:
have maximum sensitivity and biological objects give low background interference.
567:
Cy3 and Cy5 are the most popular, used typically combined for 2 colors detection.
365:
960:"Discovery of New Synthetic Dyes: Targeted Synthesis or Combinatorial Approach?"
932:
744:
332:
311:
247:
1250:
1556:
1258:
918:
860:
736:
712:
547:* Depends strongly on viscosity, temperature, and biomolecular interactions.
542:
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1029:
910:
349:
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1183:
1146:
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1055:
37:
group. Although the name derives etymologically from terms for shades of
708:
692:
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556:
377:
264:
73:
50:
34:
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532:
527:
369:
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243:
820:
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582:
560:
291:
279:
1401:
828:
287:
283:
260:
61:
46:
1333:
787:-ester) reactive group. Since NHS-esters react readily only with
672:
353:
275:
69:
20:
683:
do not have to be identical. In the dyes as used they are short
1487:
571:
295:
1490:"A nascent proteome study combining click chemistry with 2DE"
1196:
892:
869:
863:
791:
606:
327:
323:
38:
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1027:
267:
chain. Both nitrogens may each be independently part of a
99:
1313:
1028:
Ernst LA, Gupta RK, Mujumdar RB, Waggoner AS (Jan 1989).
780:
776:
88:
815:
For protein labeling, Cy3 and Cy5 dyes sometimes bear a
84:
687:
chains one or both of which ends in a highly reactive
318:
which will form an image on the film, making the film
1030:"Cyanine dye labeling reagents for sulfhydryl groups"
877:
which means a somewhat different color: "dark blue".
763:, as well as "light-up Probes," and in vivo imaging.
550:
1094:
990:
Berneth, Horst (2008). "Methine Dyes and
Pigments".
594:, this dye is also easily detected by naked eye on
1287:
1554:
1444:
1381:FluoProbes547H comparison in Confocal Microscopy
1095:Blower MD, Feric E, Weis K, Heald R (Dec 2007).
652:imaging applications, as well as the Cy7.5 dye.
1157:
1155:
993:Ullmann's Encyclopedia of Industrial Chemistry
305:
72:. Typically, they form part of a nitrogenous
68:, exactly one nitrogen atom is oxidized to an
1232:
665:
1152:
937:The IUPAC Compendium of Chemical Terminology
202:Additionally, these classes are recognized:
113:Cyanines have been classified in many ways:
79:The main application for cyanine dyes is in
33:are a synthetic dye family belonging to the
1370:FluoProbes488 comparison to FITC, Cyanine2
859:(close to "aqua") and is derived from the
585:excitation, ~570 nm emission), while
1142:
1140:
1120:
1045:
985:
983:
1391:
766:
730:
98:
1346:"Iris Dyes | Cyanine Technologies"
1233:Levitus, Marcia; Ranjit, Suman (2011).
1023:
1021:
989:
957:
342:
1555:
1137:
980:
899:European Journal of Organic Chemistry
1018:
962:. In Demchenko, Alexander P. (ed.).
958:Kim, Eunha; Park, Seung Bum (2010).
609:emission wavelengths (typically 532
810:
794:groups, which nucleic acids lack,
605:The scanners actually use diverse
13:
1290:"Cyanine Dyes (Cy2, Cy3, and Cy5)"
617:) and filter wavelengths (550-600
570:
551:Common cyanine dyes and their uses
526:Ex (nm): Excitation wavelength in
14:
1579:
1548:
1396:. Vol. 253. pp. 55–76.
966:. Berlin: Springer. p. 172.
819:group to react with amines, or a
581:fluoresces greenish yellow (~550
358:comparative genomic hybridization
314:, i.e., to increase the range of
31:tetramethylindo(di)-carbocyanines
1394:DNA Binders and Related Subjects
751:(mainly Visible range, but also
636:can replace sulfoRhodamine 101.
531:Em (nm): Emission wavelength in
41:, the cyanine family covers the
1481:
1438:
1385:
1374:
1363:
1338:
1327:
1302:
1281:
1239:Quarterly Reviews of Biophysics
1226:
1190:
1070:"Archival Lifespan of TDK CD-R"
726:
702:
1088:
1062:
951:
925:
886:
739:stained green with cyanine dye
629:Other cyanine dyes are useful:
1:
1568:Quaternary ammonium compounds
1200:Chemistry: A European Journal
1002:10.1002/14356007.a16_487.pub2
880:
850:
592:molar extinction coefficient
322:. Cyanines are also used in
94:
7:
1101:The Journal of Cell Biology
385:
306:History and use in industry
56:Chemically, cyanines are a
10:
1584:
870:
864:
666:Nomenclature and structure
18:
1251:10.1017/S0033583510000247
798:have to be modified with
368:, and various studies in
1288:Jackson ImmunoResearch.
842:Cy3 and Cy5 are used in
43:electromagnetic spectrum
19:Not to be confused with
1459:10.1002/elps.1150181133
996:. Weinheim: Wiley-VCH.
945:10.1351/goldbook.C01487
1509:10.1002/pmic.201200393
1212:10.1002/chem.200801906
1164:Bioconjugate Chemistry
1047:10.1002/cyto.990100103
911:10.1002/ejoc.200701190
823:group to react with a
759:), Microplate assays,
740:
575:
312:photographic emulsions
110:
1113:10.1083/jcb.200705163
767:Nucleic acid labeling
734:
574:
259:where two quaternary
180:Closed chain cyanines
109:III = Closed cyanines
102:
693:N-hydroxysuccinimide
675:(as shown), and the
596:electrophoresis gels
364:, which are used in
343:Use in biotechnology
105:I = Streptocyanines,
29:also referred to as
1176:10.1021/bc00020a001
122:open chain cyanines
81:biological labeling
74:heterocyclic system
66:resonance structure
60:system between two
1076:. US Digital Media
741:
576:
376:) and fluorescent
111:
107:II = Hemicyanines,
1411:978-3-540-22835-6
905:(12): 2107–2117.
524:
523:
1575:
1543:
1542:
1540:
1539:
1533:
1527:. Archived from
1494:
1485:
1479:
1478:
1442:
1436:
1435:
1429:
1425:
1423:
1415:
1389:
1383:
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1357:
1348:. Archived from
1342:
1336:
1334:SETA BioMedicals
1331:
1325:
1324:
1322:
1321:
1312:. Archived from
1306:
1300:
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1296:
1285:
1279:
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1230:
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1223:
1194:
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949:
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867:
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811:Protein labeling
709:Alexa Fluor dyes
538:Molecular weight
386:
263:are joined by a
1583:
1582:
1578:
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1573:
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1447:Electrophoresis
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1402:10.1007/b100442
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1231:
1227:
1206:(5): 1096–106.
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1145:
1138:
1093:
1089:
1079:
1077:
1068:
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1012:
988:
981:
974:
956:
952:
931:
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926:
891:
887:
883:
875:kyaneos/kyanous
853:
813:
769:
729:
717:FluoProbes dyes
705:
668:
553:
540:
535:
530:
366:transcriptomics
345:
308:
248:quinophthalones
232:
227:
222:
217:
198:
191:
174:
169:
164:
147:
142:
137:
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118:Streptocyanines
108:
106:
104:
97:
64:atoms; in each
24:
17:
12:
11:
5:
1581:
1571:
1570:
1565:
1550:
1549:External links
1547:
1545:
1544:
1480:
1453:(11): 2071–7.
1437:
1428:|journal=
1410:
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1373:
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1326:
1310:"Cyandye, LLC"
1301:
1280:
1245:(1): 123–151.
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1189:
1170:(2): 105–111.
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1107:(7): 1365–73.
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1011:978-3527306732
1010:
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933:"Cyanine dyes"
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745:Flow cytometry
737:cyanobacterium
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333:phthalocyanine
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269:heteroaromatic
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207:Neutrocyanines
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1534:on 2015-06-30
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543:Quantum yield
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103:The Cyanines:
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67:
63:
59:
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52:
48:
44:
40:
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32:
28:
22:
16:Class of dyes
1563:Cyanine dyes
1536:. Retrieved
1529:the original
1503:(1): 17–21.
1500:
1496:
1483:
1450:
1446:
1440:
1393:
1387:
1376:
1365:
1354:. Retrieved
1350:the original
1340:
1329:
1318:. Retrieved
1314:the original
1304:
1293:. Retrieved
1283:
1242:
1238:
1228:
1203:
1199:
1192:
1167:
1163:
1104:
1100:
1090:
1078:. Retrieved
1074:cdrom2go.com
1073:
1064:
1037:
1033:
991:
963:
953:
936:
927:
902:
898:
888:
874:
854:
841:
833:
817:succinimidyl
814:
775:experiments
770:
742:
727:Applications
721:
706:
703:Alternatives
669:
655:
654:
649:
645:
644:
639:
638:
633:
632:
621:and 655-695
604:
586:
578:
577:
554:
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525:
382:
378:immunoassays
346:
320:panchromatic
309:
258:
253:
240:Merocyanines
239:
229:
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206:
201:
195:
188:
179:
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161:
153:Hemicyanines
152:
144:
134:
121:
117:
112:
78:
55:
30:
26:
25:
1040:(1): 3–10.
796:nucleotides
761:Microarrays
677:side chains
557:fluorescein
435:QY 0.15 *
316:wavelengths
265:polymethine
254:Apocyanines
244:spiropyrans
35:polymethine
1557:Categories
1538:2015-06-27
1497:Proteomics
1356:2015-01-26
1320:2013-11-01
1295:2008-10-31
881:References
857:blue-green
844:proteomics
831:residues.
825:sulfhydryl
800:aminoallyl
773:microarray
749:Microscopy
660:PEGylation
561:rhodamines
533:nanometers
528:nanometers
477:(625);650
463:594;(640)
446:572;(620)
429:570;(615)
426:(512);550
370:proteomics
362:gene chips
274:, such as
242:including
224:-CN and R
58:conjugated
1430:ignored (
1420:cite book
1259:1469-8994
1034:Cytometry
919:1099-0690
851:Etymology
827:group of
821:maleimide
807:effects.
805:quenching
789:aliphatic
697:maleimide
685:aliphatic
292:quinoline
280:imidazole
261:nitrogens
187:Aryl=N=CH
160:Aryl=N=CH
95:Structure
27:Cyanines,
1517:23161590
1267:21108866
1220:19117043
1131:18166649
939:. 2014.
829:cysteine
691:such as
681:R groups
673:methines
613:and 635
520:QY 0.28
503:QY 0.28
486:QY 0.27
469:QY 0.15
452:QY 0.67
418:QY 0.12
395:Em (nm)
392:Ex (nm)
288:pyridine
284:thiazole
193:-N=Aryl
62:nitrogen
1525:9002232
1467:9420172
1275:7345293
1184:7873641
1147:CYanine
1122:2373496
1080:3 April
1056:2917472
871:κυανοῦς
865:κυάνεος
713:Dylight
650:in vivo
354:protein
302:, etc.
276:pyrrole
91:media.
70:iminium
47:near IR
21:Cyanide
1523:
1515:
1475:604007
1473:
1465:
1408:
1273:
1265:
1257:
1218:
1182:
1129:
1119:
1054:
1008:
970:
917:
689:moiety
491:Cy5.5
457:Cy3.5
389:Probe
296:indole
272:moiety
1532:(PDF)
1521:S2CID
1493:(PDF)
1471:S2CID
1271:S2CID
861:Greek
792:amine
656:Sulfo
640:Cy5.5
634:Cy3.5
607:laser
500:1128
466:1102
440:Cy3B
328:DVD-R
196:(III)
45:from
1513:PMID
1463:PMID
1432:help
1406:ISBN
1263:PMID
1255:ISSN
1216:PMID
1180:PMID
1149:dyes
1127:PMID
1082:2022
1052:PMID
1006:ISBN
968:ISBN
915:ISSN
903:2008
837:FRET
559:and
541:QY:
536:MW:
517:818
514:767
511:743
508:Cy7
497:694
494:675
483:792
480:670
474:Cy5
460:581
449:658
443:558
432:767
423:Cy3
415:714
412:506
409:489
406:Cy2
374:FRET
360:and
335:and
326:and
324:CD-R
246:and
234:-CHO
228:N=CH
218:N=CH
172:(II)
133:N=CH
87:and
39:blue
1505:doi
1455:doi
1398:doi
1247:doi
1208:doi
1172:doi
1117:PMC
1109:doi
1105:179
1042:doi
998:doi
941:doi
907:doi
785:NHS
781:RNA
779:or
777:DNA
771:In
695:or
646:Cy7
587:Cy5
579:Cy3
398:MW
352:or
337:azo
166:-NR
145:(I)
139:-NR
120:or
89:DVD
49:to
1559::
1519:.
1511:.
1501:13
1499:.
1495:.
1469:.
1461:.
1451:18
1449:.
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1422:}}
1418:{{
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1261:.
1253:.
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1241:.
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1178:.
1166:.
1154:^
1139:^
1125:.
1115:.
1103:.
1099:.
1072:.
1050:.
1038:10
1036:.
1032:.
1020:^
1004:.
982:^
935:.
913:.
901:.
897:.
757:IR
755:,
753:UV
747:,
735:A
715:,
711:,
699:.
623:nm
619:nm
615:nm
611:nm
583:nm
563:.
380:.
339:.
298:,
294:,
290:,
286:,
282:,
278:,
85:CD
76:.
53:.
51:UV
1541:.
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1477:.
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1359:.
1323:.
1298:.
1277:.
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1222:.
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1186:.
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1168:4
1133:.
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1084:.
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1014:.
1000::
976:.
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921:.
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868:/
250:.
231:n
226:2
221:n
216:2
214:R
209::
190:n
182::
168:2
163:n
155::
141:2
136:n
131:2
129:R
124::
23:.
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