ADN/RNA sekans -pacbio sekansyèl
Karakteristik
De mòd sekans sou Pacbio Sequencer: Kontini Long Li (CLR) ak konsansis sikilè li (CCS)
| Mòd sekans | Gwosè bibliyotèk | Done teyorikSede (pou chak selil) | Single-bazPresizyon | Aplikasyon |
| Clr | 20kb, 30kb, elatriye. | 80 GB a 130 GB | Approx. 85% | De Novo, Sv rele, elatriye. |
| CCS | 15-20 KB | 14 a 40 GB/selil (fen II) 70 a 110 GB/selil (Revio) Depann sou echantiyon yo | Approx. 99% | De Novo, SNP/Indel/SV rele, ISO-seq, |
Konparezon nan pèfòmans lan ak karakteristik nan Revio ak fen II
| Tèm | Fen II Sistèm | Sistèm Revio | Ogmantasyon |
| Pi wo dansite | 8 milyon ZMWs | 25 milyon ZMWs | 3x |
| Etap endepandan | 1 | 4 | 4x |
| Pi kout fwa kouri | 30 èdtan | 24 èdtan | 1.25x |
| 30x hifi jnom imen / ane | 88 | 1,300 | 15x an jeneral |
Avantaj Sèvis
● Plis pase 8 ane eksperyans sou Pacbio sekans platfòm ak dè milye de pwojè fèmen ak divès kalite espès.
● Fully ekipe ak dènye platfòm yo sekans Pacbio, Revio garanti ase sekans debi.
● Pi vit vire-alantou tan, pi wo done sede ak done pi egzat.
● Kontribye nan dè santèn de gwo enpak piblikasyon ki baze sou Pacbio.
Egzanp kondisyon
| Egzanp Kalite | Montan | Konsantrasyon （qubit®） | Volim | Pite | Lòt lòt |
| ADN jenomik | Depann sou kondisyon done | ≥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 yo dwe mezire pa qubit ak qubit/nanopore = 0.8-2.5 |
| Total RNA | ≥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
Syen
Done analiz
Livrezon Pwojè
1.in-kay done sede
Done ki soti nan 63 selil CCS (ki soti nan 26 espès)
| Done-Pacbio-CCS-15 KB | Mwayèn | Max | Min | Medyàn |
| 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 |
| Subreads 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 | Medyàn |
| Sede - subreads (GB) | 142.20 | 291.40 | 50.55 | 142.49 |
| Polymerase N50 | 39,456 | 121,191 | 15,389 | 35,231 |
| Subreads 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 done sede
| Echantiyon | CCS li num | Total baz CCS (BP) | CCS li N50 (BP) | CCS vle di longè (BP) | CCS pi long li (BP) | Subreads baz (BP) | CCS to (%) |
| Pb_bmkxxx | 3,444,159 | 54,164,122,586 | 15,728 | 15,726 | 36,110 | 863,326,330,465 | 6.27 |
