ADN/RNA sekans -PacBio Sequencer
Karakteristik
De mòd sekans sou sekans PacBio: Continuous Long Read (CLR) ak Circular Consensus Read (CCS)
| Sekans mòd | Gwosè Bibliyotèk | Done teyorikSede (pa selil) | Single-bazPresizyon | Aplikasyon |
| CLR | 20Kb, 30Kb, elatriye. | 80 Gb pou 130 Gb | Apeprè.85% | De novo, SV rele, elatriye. |
| CCS | 15-20 Kb | 14 a 40 Gb/Selil (Sekil II) 70 a 110 Gb/Selil (Revio) Sa depann de echantiyon yo | Apeprè.99% | De novo, SNP/Indel/SV rele, Iso-Seq, |
Konparezon pèfòmans ak karakteristik Revio ak Sequel II
| Regleman | Sequel II sistèm | Revio sistèm | Ogmante |
| Pi wo dansite | 8 milyon ZMWs | 25 milyon ZMWs | 3x |
| Etap endepandan | 1 | 4 | 4x |
| Pi kout tan kouri | 30 èdtan | 24 èdtan | 1.25x |
| 30X HiFi jenom imen / ane | 88 | 1,300 | 15x an jeneral |
Avantaj sèvis yo
● Plis pase 8 ane eksperyans sou platfòm sekans PacBio ak plizyè milye pwojè fèmen ak divès espès.
● Konplètman ekipe ak dènye platfòm PacBio Sekans, Revio pou garanti ase debi sekans.
● Pi vit vire tan, pi wo sede done ak done ki pi egzak.
● Kontribye nan plizyè santèn piblikasyon ki baze sou PacBio ki gen gwo enpak.
Egzanp kondisyon yo
| Kalite echantiyon | Kantite lajan | Konsantrasyon （Qubit®） | Volim | Pite | Lòt moun |
| ADN jenomik | Depann sou Done Egzijans | ≥50 ng/μl | ≥15μl | OD260/280 = 1.7-2.2; OD260/230 = 1.8-2.5; Klè pik nan 260 nm,pa gen kontaminasyon | Konsantrasyon bezwen mezire pa Qubit ak Qubit/Nanopore = 0.8-2.5 |
| RNA total | ≥1.2μg | ≥120 ng/μl | ≥15μl | OD260/280 = 1.7-2.5; OD260/230 = 0.5-2.5;pa gen kontaminasyon | Valè RIN ≥7.5 5≥28S/18S≥1 |
Sèvis Workflow
Preparasyon echantiyon
Konstriksyon bibliyotèk
Sekans
Analiz done
Livrezon pwojè
1.In-house Done Sede
Done ki soti nan 63 selil CCS (ki soti nan 26 espès)
| DONE-PacBio-CCS-15 Kb | Mwayèn | Max | Min | Mwayèn |
| Sede - subreads (Gb) | 421.12 | 544.27 | 221.38 | 426.58 |
| Yiled - CCS (Gb) | 25.93 | 38.59 | 10.86 | 25.43 |
| Polymerase N50 | 145,651 | 175,430 | 118,118 | 144,689 |
| Subread N50 | 17,509 | 23,924 | 12,485 | 17,584 |
| CCS N50 | 14,490 | 19,034 | 9,876 | 14,747 |
| Mwayèn longè-Polymerase | 67,995 | 89,379 | 49,664 | 66,433 |
| Mwayèn longè-Subreads | 15,866 | 21,036 | 11,657 | 16,012 |
| Mwayèn longè-CCS | 14,489 | 19,074 | 8,575 | 14,655 |
Done ki soti nan 16 selil CLR (ki soti nan 76 espès)
| DONE-PacBio-CLR-30Kb | Mwayèn | Max | Min | Mwayèn |
| Sede - subreads (Gb) | 142.20 | 291.40 | 50.55 | 142.49 |
| Polymerase N50 | 39,456 | 121,191 | 15,389 | 35,231 |
| Subread N50 | 28,490 | 41,012 | 14,430 | 29,063 |
| Mwayèn longè-Polymerase | 22,063 | 48,886 | 8,747 | 21,555 |
| Mwayèn longè-Subreads | 17,720 | 27,225 | 8,293 | 17,779 |
2.Data QC - DemoEstatistik sou rannman done
| Egzanp | ccs Li Num | Total ccs baz (bp) | ccs Li N50 (bp) | ccs Longè Mwayèn (bp) | ccs pi long lekti (bp) | subli baz (bp) | pousantaj ccs (%) |
| PB_BMKxxx | 3,444,159 | 54,164,122,586 | 15,728 | 15,726 | 36,110 | 863,326,330,465 | 6.27 |
