266:
366:(containing sulfurylase and luciferase) onto a PicoTiterPlate device. The device was centrifuged to deposit the beads into the wells. The layer of Enzyme Beads ensured that the DNA beads remained positioned in the wells during the sequencing reaction. The bead-deposition process was designed to maximize the number of wells that contain a single amplified library bead.
384:. The signal strength was proportional to the number of nucleotides; for example, homopolymer stretches, incorporated in a single nucleotide flow, generated a greater signal than single nucleotides. However, the signal strength for homopolymer stretches was linear only up to eight consecutive nucleotides, after which the signal fell off rapidly. Data were stored in
379:
to the template strand was added into a well, the polymerase extended the existing DNA strand by adding nucleotide(s). Addition of one (or more) nucleotide(s) generated a light signal that was recorded by the CCD camera in the instrument. This technique was based on sequencing-by-synthesis and called
239:
Genomic DNA was fractionated into smaller fragments (300-800 base pairs) and polished (made blunt at each end). Short adaptors were then ligated onto the ends of the fragments. These adaptors provided priming sequences for both amplification and sequencing of the sample-library fragments. One adaptor
619:
Wheeler, D. A.; Srinivasan, M.; Egholm, M.; Shen, Y.; Chen, L.; McGuire, A.; He, W.; Chen, Y. J.; Makhijani, V.; Roth, G. T.; Gomes, X.; Tartaro, K.; Niazi, F.; Turcotte, C. L.; Irzyk, G. P.; Lupski, J. R.; Chinault, C.; Song, X.-Z.; Liu, Y.; Yuan, Y.; Nazareth, L.; Qin, X.; Muzny, D. M.; Margulies,
251:
The sstDNA library was immobilized onto beads. The beads containing a library fragment carried a single sstDNA molecule. The bead-bound library was emulsified with the amplification reagents in a water-in-oil mixture. Each bead was captured within its own microreactor where PCR amplification occurs.
230:
on the market. In 2008, 454 Sequencing launched the GS FLX Titanium series reagents for use on the Genome
Sequencer FLX instrument, with the ability to sequence 400-600 million base pairs per run with 400-500 base pair read lengths. In late 2009, 454 Life Sciences introduced the GS Junior System, a
134:
and was originally known as 454 Corporation, a subsidiary of CuraGen. For their method for low-cost gene sequencing, 454 Life
Sciences was awarded the Wall Street Journal's Gold Medal for Innovation in the Biotech-Medical category in 2005. The name 454 was the code name by which the project was
248:-coated beads. After nick repair, the non-biotinylated strand was released and used as a single-stranded template DNA (sstDNA) library. The sstDNA library was assessed for its quality, and the optimal amount (DNA copies per bead) needed for emPCR is determined by titration.
374:
were added sequentially in a fixed order across the PicoTiterPlate device during a sequencing run. During the nucleotide flow, millions of copies of DNA bound to each of the beads were sequenced in parallel. When a nucleotide
369:
The loaded PicoTiterPlate device were placed into the Genome
Sequencer FLX Instrument. The fluidics sub-system delivered sequencing reagents (containing buffers and nucleotides) across the wells of the plate. The four DNA
511:
Green, Richard E.; Krause, Johannes; Ptak, Susan E.; Briggs, Adrian W.; Ronan, Michael T.; Simons, Jan F.; Du, Lei; Egholm, Michael; Rothberg, Jonathan M.; Paunovic, Maja; Pääbo, Svante (November 2006).
161:
acquired 454 Life
Sciences for US$ 154.9 million. It remained a separate business unit. In October 2013, Roche announced that it would shut down 454, and stop supporting the platform by mid-2016.
462:
937:
932:
947:
962:
942:
605:
669:
417:
436:
737:"454 Life Sciences Unveils New Bench Top Sequencer, Significant Improvements to the Genome Sequencer FLX System Including 1,000 bp Reads for 2010"
913:
180:
system capable of sequencing roughly 400-600 megabases of DNA per 10-hour run on the Genome
Sequencer FLX with GS FLX Titanium series reagents.
972:
736:
967:
362:
Single-stranded template DNA library beads were added to the DNA Bead
Incubation Mix (containing DNA polymerase) and were layered with
493:
470:
330:
122:
in 2007 and shut down by Roche in 2013 when its technology became noncompetitive, although production continued until mid-2016.
302:
376:
309:
283:
561:
349:
17:
316:
587:
287:
298:
673:
957:
219:
was also packed into the well. The PicoTiterPlate was then placed into the GS FLX System for sequencing.
227:
151:
192:
740:
385:
952:
276:
138:
In
November 2006, Rothberg, Michael Egholm, and colleagues at 454 published a cover article with
562:"Roche - Roche acquires 454 Life Sciences to strengthen presence in ultra-fast gene sequencing"
323:
606:"Following Roche's Decision to Shut Down 454, Customers Make Plans to Move to Other Platforms"
907:
119:
111:
72:
60:
869:
764:"Titration-free massively parallel pyrosequencing using trace amounts of starting material"
633:
525:
8:
164:
In May 2007, 454 published the results of
Project "Jim": the sequencing of the genome of
873:
637:
529:
890:
857:
833:
812:
788:
763:
223:
895:
838:
793:
718:
651:
543:
158:
139:
131:
885:
877:
828:
820:
783:
775:
708:
641:
533:
143:
856:
Margulies, Marcel; Michael Egholm; 54 additional coauthors (September 15, 2005).
713:
696:
437:"Quantum-Si taps founder Rothberg, prolific medtech entrepreneur, as interim CEO"
212:
165:
397:
381:
208:
196:
187:
and adapter-ligated DNA fragments to small DNA-capture beads in a water-in-oil
177:
115:
855:
926:
87:
41:
899:
858:"Genome Sequencing in Open Microfabricated High Density Picoliter Reactors"
842:
797:
722:
655:
622:"The complete genome of an individual by massively parallel DNA sequencing"
547:
245:
565:
779:
200:
881:
646:
621:
538:
513:
371:
216:
135:
referred to at CuraGen, and the numbers have no known special meaning.
418:"Roche to close 454 Life Sciences as it reduces gene sequencing focus"
739:(Press release). 454 Life Sciences. November 19, 2009. Archived from
252:
This resulted in bead-immobilized, clonally amplified DNA fragments.
184:
147:
265:
188:
63:
in 2007 and shut down by Roche in 2013 (production ceased mid-2016)
204:
91:
824:
697:"Next Generation Sequencing: From Basic Research to Diagnostics"
241:
620:
M.; Weinstock, G. M.; Gibbs, R. A.; Rothberg, J. M. (2008).
154:
to complete the sequence of the
Neanderthal genome by 2009.
618:
813:"Pyrosequencing: a simple method for accurate genotyping"
195:. Each DNA-bound bead was placed into a ~29 ÎĽm well on a
514:"Analysis of one million base pairs of Neanderthal DNA"
231:
bench top version of the Genome
Sequencer FLX System.
510:
191:. The DNA fixed to these beads was then amplified by
938:
Defunct manufacturing companies based in Connecticut
933:
Defunct biotechnology companies of the United States
810:
234:
670:"Project Jim: Watson's Personal Genome Goes Public"
290:. Unsourced material may be challenged and removed.
494:"Company Says It Mapped Genes of Virus in One Day"
948:Companies based in New Haven County, Connecticut
924:
244:tag for immobilization of the DNA library onto
963:Biotechnology companies disestablished in 2013
588:"Roche snares 454's sequencing tech in buyout"
849:
504:
150:of the Neanderthal genome, and initiated the
943:Biotechnology companies established in 2000
912:: CS1 maint: numeric names: authors list (
176:454 Sequencing used a large-scale parallel
868:(7057). Nature Publishing Group: 376–380.
889:
832:
787:
712:
645:
537:
350:Learn how and when to remove this message
168:, co-discoverer of the structure of DNA.
491:
415:
14:
925:
973:2013 disestablishments in Connecticut
761:
460:
388:(SFF) files for downstream analysis.
110:was a biotechnology company based in
694:
434:
288:adding citations to reliable sources
259:
114:that specialized in high-throughput
492:Pollack, Andrew (August 22, 2003).
461:Totty, Michael (October 24, 2005).
24:
968:2000 establishments in Connecticut
811:King, C; Scott-Horton, T. (2008).
435:Park, Andrea (February 15, 2022).
416:Hollmer, Mark (October 17, 2013).
25:
984:
235:DNA library preparation and emPCR
130:454 Life Sciences was founded by
264:
804:
755:
729:
688:
662:
275:needs additional citations for
762:Zheng, Z; et al. (2010).
612:
598:
580:
554:
485:
454:
428:
409:
13:
1:
695:Karl, V; et al. (2009).
255:
228:next-generation DNA sequencer
171:
146:describing the first million
101:Sequencing of genetic samples
714:10.1373/clinchem.2008.112789
183:The system relied on fixing
7:
391:
240:(Adaptor B) contained a 5'-
10:
989:
152:Neanderthal Genome Project
125:
97:
83:
67:
55:
47:
37:
403:
386:standard flowgram format
467:The Wall Street Journal
222:454 released the GS20
473:on September 30, 2015
118:. It was acquired by
112:Branford, Connecticut
73:Branford, Connecticut
27:Biotechnology company
743:on November 15, 2011
676:on November 21, 2008
284:improve this article
157:In late March 2007,
882:10.1038/nature03959
874:2005Natur.437..376M
647:10.1038/nature06884
638:2008Natur.452..872W
608:. October 22, 2013.
539:10.1038/nature05336
530:2006Natur.444..330G
299:"454 Life Sciences"
226:in 2005, the first
34:
958:Genomics companies
780:10.1093/nar/gkq332
701:Clinical Chemistry
568:on August 29, 2012
498:The New York Times
224:sequencing machine
32:
768:Nucleic Acids Res
632:(7189): 872–876.
594:. March 28, 2007.
524:(7117): 330–336.
360:
359:
352:
334:
159:Roche Diagnostics
132:Jonathan Rothberg
108:454 Life Sciences
105:
104:
88:Genome sequencers
33:454 Life Sciences
18:454 life sciences
16:(Redirected from
980:
918:
917:
911:
903:
893:
853:
847:
846:
836:
808:
802:
801:
791:
759:
753:
752:
750:
748:
733:
727:
726:
716:
692:
686:
685:
683:
681:
672:. Archived from
666:
660:
659:
649:
616:
610:
609:
602:
596:
595:
584:
578:
577:
575:
573:
564:. Archived from
558:
552:
551:
541:
508:
502:
501:
489:
483:
482:
480:
478:
469:. Archived from
458:
452:
451:
449:
447:
432:
426:
425:
413:
355:
348:
344:
341:
335:
333:
292:
268:
260:
35:
31:
21:
988:
987:
983:
982:
981:
979:
978:
977:
923:
922:
921:
905:
904:
854:
850:
809:
805:
760:
756:
746:
744:
735:
734:
730:
693:
689:
679:
677:
668:
667:
663:
617:
613:
604:
603:
599:
586:
585:
581:
571:
569:
560:
559:
555:
509:
505:
490:
486:
476:
474:
463:"A Better Idea"
459:
455:
445:
443:
433:
429:
414:
410:
406:
394:
356:
345:
339:
336:
293:
291:
281:
269:
258:
237:
213:ATP sulfurylase
203:chip. A mix of
174:
128:
79:
75:
28:
23:
22:
15:
12:
11:
5:
986:
976:
975:
970:
965:
960:
955:
953:DNA sequencing
950:
945:
940:
935:
920:
919:
848:
803:
754:
728:
687:
661:
611:
597:
579:
553:
503:
484:
453:
441:Fierce Biotech
427:
422:Fierce Biotech
407:
405:
402:
401:
400:
398:DNA Sequencing
393:
390:
382:pyrosequencing
358:
357:
272:
270:
263:
257:
254:
236:
233:
209:DNA polymerase
197:PicoTiterPlate
178:pyrosequencing
173:
170:
127:
124:
116:DNA sequencing
103:
102:
99:
95:
94:
85:
81:
80:
77:
71:
69:
65:
64:
57:
53:
52:
49:
45:
44:
39:
26:
9:
6:
4:
3:
2:
985:
974:
971:
969:
966:
964:
961:
959:
956:
954:
951:
949:
946:
944:
941:
939:
936:
934:
931:
930:
928:
915:
909:
901:
897:
892:
887:
883:
879:
875:
871:
867:
863:
859:
852:
844:
840:
835:
830:
826:
822:
818:
814:
807:
799:
795:
790:
785:
781:
777:
773:
769:
765:
758:
742:
738:
732:
724:
720:
715:
710:
706:
702:
698:
691:
675:
671:
665:
657:
653:
648:
643:
639:
635:
631:
627:
623:
615:
607:
601:
593:
592:FierceBiotech
589:
583:
567:
563:
557:
549:
545:
540:
535:
531:
527:
523:
519:
515:
507:
499:
495:
488:
472:
468:
464:
457:
442:
438:
431:
423:
419:
412:
408:
399:
396:
395:
389:
387:
383:
378:
377:complementary
373:
367:
365:
354:
351:
343:
332:
329:
325:
322:
318:
315:
311:
308:
304:
301: –
300:
296:
295:Find sources:
289:
285:
279:
278:
273:This article
271:
267:
262:
261:
253:
249:
247:
243:
232:
229:
225:
220:
218:
214:
210:
206:
202:
198:
194:
190:
186:
181:
179:
169:
167:
162:
160:
155:
153:
149:
145:
141:
136:
133:
123:
121:
117:
113:
109:
100:
96:
93:
89:
86:
82:
74:
70:
66:
62:
58:
54:
50:
46:
43:
42:Biotechnology
40:
36:
30:
19:
908:cite journal
865:
861:
851:
816:
806:
774:(13): e137.
771:
767:
757:
747:November 19,
745:. Retrieved
741:the original
731:
707:(4): 41–47.
704:
700:
690:
678:. Retrieved
674:the original
664:
629:
625:
614:
600:
591:
582:
572:November 14,
570:. Retrieved
566:the original
556:
521:
517:
506:
497:
487:
475:. Retrieved
471:the original
466:
456:
446:February 22,
444:. Retrieved
440:
430:
421:
411:
368:
364:Enzyme Beads
363:
361:
346:
337:
327:
320:
313:
306:
294:
282:Please help
277:verification
274:
250:
246:streptavidin
238:
221:
182:
175:
166:James Watson
163:
156:
140:Svante Pääbo
137:
129:
107:
106:
68:Headquarters
59:Acquired by
29:
825:10.3791/630
372:nucleotides
340:August 2022
201:fiber optic
927:Categories
310:newspapers
256:Sequencing
217:luciferase
172:Technology
148:base pairs
817:J Vis Exp
477:March 13,
185:nebulized
900:16056220
843:19066560
798:20435675
723:19246620
680:June 16,
656:18421352
548:17108958
392:See also
207:such as
189:emulsion
98:Services
92:reagents
84:Products
38:Industry
891:1464427
870:Bibcode
834:2582836
789:2910068
634:Bibcode
526:Bibcode
324:scholar
205:enzymes
126:History
48:Founded
898:
888:
862:Nature
841:
831:
819:(11).
796:
786:
721:
654:
626:Nature
546:
518:Nature
326:
319:
312:
305:
297:
242:biotin
215:, and
144:Nature
404:Notes
331:JSTOR
317:books
120:Roche
61:Roche
914:link
896:PMID
839:PMID
794:PMID
749:2009
719:PMID
682:2007
652:PMID
574:2012
544:PMID
479:2017
448:2022
303:news
199:, a
56:Fate
51:2000
886:PMC
878:doi
866:437
829:PMC
821:doi
784:PMC
776:doi
709:doi
642:doi
630:452
534:doi
522:444
286:by
193:PCR
142:in
78:USA
929::
910:}}
906:{{
894:.
884:.
876:.
864:.
860:.
837:.
827:.
815:.
792:.
782:.
772:38
770:.
766:.
717:.
705:55
703:.
699:.
650:.
640:.
628:.
624:.
590:.
542:.
532:.
520:.
516:.
496:.
465:.
439:.
420:.
211:,
90:,
76:,
916:)
902:.
880::
872::
845:.
823::
800:.
778::
751:.
725:.
711::
684:.
658:.
644::
636::
576:.
550:.
536::
528::
500:.
481:.
450:.
424:.
353:)
347:(
342:)
338:(
328:·
321:·
314:·
307:·
280:.
20:)
Text is available under the Creative Commons Attribution-ShareAlike License. Additional terms may apply.