Taxanaha DNA/RNA -PacBio Sequencer
Astaamaha
Laba hab oo isku xigxiga oo ku saabsan taxanaha PacBio: Akhriska dheer ee Joogtada ah (CLR) iyo Akhriska Oggolaanshaha Wareegtada (CCS)
| Habka isku xigxiga | Cabbirka Maktabadda | Xogta AragtidaWax-soosaarka (Qofkii unug) | Hal-saldhigSaxnaanta | Codsiyada |
| CLR | 20Kb, 30Kb, iwm. | 80 Gb ilaa 130 Gb | Qiyaastii85% | De novo, wicitaanka SV, iwm. |
| CCS | 15-20 Kb | 14 ilaa 40 Gb/Ceel (Taxanaha II) 70 ilaa 110 Gb/Ceel (Revio) Waxay kuxirantahay shaybaarada | Qiyaastii99% | De novo, SNP/Indel/SV wacitaanka, Iso-Seq, |
Isbarbardhigga waxqabadka iyo astaamaha Revio iyo Taxanaha II
| Shuruudaha | Taxanaha II nidaamka | Nidaamka Revio | Kordhiyo |
| Cufnaanta sare | 8 milyan oo ZMW | 25 milyan oo ZMW | 3x |
| Marxalado madaxbannaan | 1 | 4 | 4x |
| Waqtiyada orodka gaaban | 30 saacadood | 24 saacadood | 1.25x |
| 30X HiFi hiddo-wadaha aadanaha / sanadka | 88 | 1,300 | 15x guud ahaan |
Faa'iidooyinka Adeegga
● In ka badan 8 sano oo waayo-aragnimo ah oo ku saabsan madal taxane ah oo PacBio ah oo leh kumanaan mashruuc oo xiran oo leh noocyo kala duwan.
● Si buuxda ugu qalabaysan aaladaha Taxanaha PacBio ee ugu dambeeyay, Revio si loo dammaanad qaado soo saarista isku xigxiga ee ku filan.
● Wakhtiga ka-wareejinta degdega ah, xog-soosaarka sare iyo xog sax ah.
● Wax ku biiriyay boqolaal daabacado ku salaysan saamaynta sare leh ee PacBio.
Shuruudaha Tusaalaha
| Tusaalaha Nooca | Qadarka | Xooga saarida (Qubit®) | Mugga | daahirnimo | Kuwo kale |
| DNA-da genomiga ah | Waxay kuxirantahay Shuruudaha Xogta | ≥50 ng/μl | ≥15ml | OD260/280=1.7-2.2; OD260/230=1.8-2.5; Nadiifi ugu sarreeya 260 nmwasakhaysan | Xoog saarista waxa loo baahan yahay in lagu cabbiro Qubit iyo Qubit/Nanopore = 0.8-2.5 |
| Wadarta RNA | ≥1.2μg | ≥120 ng/μl | ≥15ml | OD260/280=1.7-2.5; OD260/230=0.5-2.5;wasakhaysan | Qiimaha RIN ≥7.5 5≥28S/18S≥1 |
Socodka Shaqada Adeegga
Tusaalaha diyaarinta
Dhismaha maktabadda
Isku xigxiga
Falanqaynta xogta
gaarsiinta mashruuca
1.In-guriga Xog-Soosaarka
Xog laga soo saaray 63 unug CCS (oo ka yimid 26 nooc)
| XOGTA-PacBio-CCS-15 Kb | Celcelis ahaan | Max | Min | Dhexdhexaad |
| Wax-soo-saar-hoosaadyo (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 |
| Hoos ka akhriso N50 | 17,509 | 23,924 | 12,485 | 17,584 |
| CCS N50 | 14,490 | 19,034 | 9,876 | 14,747 |
| Celceliska dhererka-Polymerase | 67,995 | 89,379 | 49,664 | 66,433 |
| Dhererka celceliska-Subroodhida | 15,866 | 21,036 | 11,657 | 16,012 |
| Celceliska dhererka-CCS | 14,489 | 19,074 | 8,575 | 14,655 |
Xog laga soo saaray 16 unug CLR (laga bilaabo 76 nooc)
| DATA-PacBio-CLR-30Kb | Celcelis ahaan | Max | Min | Dhexdhexaad |
| Wax-soo-saar-hoosaadyo (Gb) | 142.20 | 291.40 | 50.55 | 142.49 |
| Polymerase N50 | 39,456 | 121,191 | 15,389 | 35,231 |
| Hoos ka akhriso N50 | 28,490 | 41,012 | 14,430 | 29,063 |
| Celceliska dhererka-Polymerase | 22,063 | 48,886 | 8,747 | 21,555 |
| Dhererka celceliska-Subroodhida | 17,720 | 27,225 | 8,293 | 17,779 |
2.Data QC - DemoTirakoobka wax-soo-saarka xogta
| Tusaale | ccs Akhri tir | Wadarta saldhigyada ccs (bp) | ccs Wuxuu akhriyaa N50 (bp) | Dhererka celceliska ccs (bp) | Akhriska ugu Dheer (bp) | Saldhigyada hoose (bp) | heerka ccs (%) |
| PB_BMKxxx | 3,444,159 | 54,164,122,586 | 15,728 | 15,726 | 36,110 | 863,326,330,465 | 6.27 |
