Dna / rna sequencing -pacbio sequencer
Zvimiro
Miviri yekutsvagisa modes paPacbio Sequencer: inoenderera mberi kuverenga kuverenga (clr) uye kutenderera kubvumirana kuverenga (CCS)
| Sequencing Mode | Saizi raibhurari | Theoretical dataGoho (pasero) | Imwe-baseKunyatso | Zvishandiso |
| CLR | 20kb, 30kb, nezvimwewo | 80 GB kusvika 130 GB | Approx. 85% | De novo, Sv Kudana, Etc. |
| CCC | 15-20 KB | 14 kusvika 40 GB / Sero (Sequel II) 70 kusvika 110 GB / Sero (revio) Zvinoenderana nesamples | Approx. 99% | De novo, SnP / Indel / SV Kufona, iso-seq, |
Kuenzanisa kwekuita uye maficha eRevio uye Sequel II
| Mazwi | Sequel II system | Revio System | Kuwedzera |
| Yakakwira density | Mamirioni masere ZMMS | Mamirioni makumi maviri nemashanu ZMMS | 3x |
| Matanho akazvimirira | 1 | 4 | 4x |
| Pfupi pfupi nguva | Maawa makumi matatu | Maawa makumi maviri nemana | 1.25x |
| 30x hifi vanhu / gore | 88 | 1,300 | 15x zvese |
Zvigadzirwa zvebasa
● Anopfuura makore masere anowana paPacbio Sequencing chikuva nezviuru zvemapurojekiti akavharwa nemarudzi akasiyana siyana.
● Zvakarongedzwa zvizere neChikamu cheCacbio Sequencing mapuratifomu, revio to laventee yakakwana kutevedzana yekupfuura.
● Kukurumidza kutendeuka-kutenderedza nguva, yepamusoro data goho uye data rakaringana.
● Akapatsanurwa mumazana emabhuku eCacbio-akavhomora.
Zvinodiwa Muenzaniso
| Sampuli mhando | Huwandu | Concentration (Qubit®) | Vhoriyamu | Kuchena | Vamwe |
| Genomic DNA | Zvinoenderana nedata zvinodiwa | ≥50 NG / μL | ≥15μ | Od260 / 280 = 1,7-2.2; Od260 / 230 = 1.8-2.5; Bvisa peak pa 260 nm,Hapana Kusvibisa | Kusimbisa kunoda kuyerwa ne quibit uye quibit / nanopore = 0.8-2.5 |
| Yakazara rna | ≥1.2μG | ≥120 NG / μL | ≥15μ | Od260 / 280 = 1,7.5; Od260 / 230 = 0.5-2.5;Hapana Kusvibisa | Rin Kukosha ≥7.5 5828s / 18s≥1 |
Basa rekushandira
Kuenzanisira Kugadzirira
Raibhurari Kuvaka
Kutevedzana
Kuongororwa kwedata
Kutumirwa kwepurojekiti
1.In-imba data goho
Data yakagadzirwa kubva pamakumi matanhatu eCCS cell (kubva ku26 marudzi)
| Data-Pacbio-CCS-15 KB | Pakati | Max | Min | Pakati |
| Goho - rugwaro rwokutanga (GB) | 421.12 | 544.27 | 221.38 | 426.58 |
| Yadhi - CCS (GB) | 25.93 | 38.59 | 10.86 | 25.43 |
| Polymerase N50 | 145,651 | 175,430 | 118,118 | 144,689 |
| Zvikungo N50 | 17,509 | 23,924 | 12,485 | 17,584 |
| CCC N50 | 14,490 | 19,034 | 9,876 | 14,747 |
| Avhareji kureba-polymerase | 67,995 | 89,379 | 49,684 | 66,433 |
| Avhareji kureba-kukurumidza | 15,866 | 21,036 | 11,657 | 16,012 |
| Avhareji kureba-ccs | 14,489 | 19,074 | 8,575 | 14,655 |
Data yakagadzirwa kubva 16 crr masero (kubva pamarudzi manomwe)
| Data-pacbio-clr-30kb | Pakati | Max | Min | Pakati |
| Goho - rugwaro rwokutanga (GB) | 142.20 | 291.40 | 50.55 | 142.49 |
| Polymerase N50 | 39,45 49,49 | 121,191 | 15,389 | 35,231 |
| Zvikungo N50 | 28,490 | 41,012 | 14,430 | 29,063 |
| Avhareji kureba-polymerase | 22,063 | 48,886 | 8,747 | 21,555 |
| Avhareji kureba-kukurumidza | 17,720 | 27,225 | 8,293 | 17,779 |
2.data QC - DemoStatistics pane data goho
| Sampuli | CCS anoverenga nhamba | Yakazara CCs zvigadziko (BP) | CCS anoverenga N50 (BP) | CCC zvinoreva kureba (BP) | CCC kureba kwazvo kuverenga (BP) | Zvinyorwa Zvekutanga (BP) | CCC rate (%) |
| Pb_bmkxxx | 3,444,159 | 54,164,12,586 | 15,728 | 15,726 | 36,110 | 863,326,3,465 | 6.27 |
