DNA / RNA sequencing -PacBio Sequencer
Features
Twa sequencing-modi op PacBio sequencer: Continuous Long Read (CLR) en Circular Consensus Read (CCS)
| Sequencing Mode | Biblioteek Grutte | Teoretyske gegevensOpbringst (per sel) | Single-baseKrektens | Oanfraach |
| CLR | 20Kb, 30Kb, ensfh. | 80 Gb oant 130 Gb | Ca.85% | De novo, SV calling, ensfh. |
| CCS | 15-20 Kb | 14 oant 40 Gb/sel (ferfolch II) 70 oant 110 Gb / sel (Revio) Hinget ôf fan de samples | Ca.99% | De novo, SNP/Indel/SV calling, Iso-Seq, |
Fergeliking fan 'e prestaasjes en funksjes fan Revio en Sequel II
| Betingsten | Sequel II systeem | Revio systeem | Tanimme |
| Hegere tichtheid | 8 miljoen ZMWs | 25 miljoen ZMWs | 3x |
| Unôfhinklike stadia | 1 | 4 | 4x |
| Koartere run tiden | 30 oeren | 24 oeren | 1,25x |
| 30X HiFi minsklike genomen / jier | 88 | 1.300 | 15x oer |
Service Foardielen
● Mear dan 8 jier ûnderfining op PacBio sequencing platfoarm mei tûzenen sletten projekten mei ferskate soarten.
● Folslein ynrjochte mei de lêste PacBio Sequencing-platfoarms, Revio om genôch sequencing-trochput te garandearjen.
● Faster turn-around tiid, hegere gegevens opbringst en krekter gegevens.
● Bydroegen yn hûnderten pacBio-basearre publikaasjes mei hege ynfloed.
Sample Requirements
| Sample Type | Tal | Konsintraasje (Qubit®) | Folume | Purity | Oaren |
| Genomysk DNA | Ofhinklik fan Data Requirement | ≥50 ng/μl | ≥15μl | OD260/280=1,7-2,2; OD260/230=1.8-2.5; Dúdlike pyk by 260 nm,gjin fersmoargingen | Konsintraasje moat wurde mjitten troch Qubit en Qubit / Nanopore = 0.8-2.5 |
| Totaal RNA | ≥1.2μg | ≥120 ng/μl | ≥15μl | OD260/280=1,7-2,5; OD260/230=0.5-2.5;gjin fersmoargingen | RIN wearde ≥7,5 5≥28S/18S≥1 |
Service Workflow
Sample tarieding
Biblioteekbou
Sequencing
Data analyze
Projekt levering
1.In-house Data Yield
Gegevens oanmakke út 63 CCS-sellen (fan 26 soarten)
| DATA-PacBio-CCS-15 Kb | Trochsneed | Max | Min | Mediaan |
| Yield - 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 |
| Gemiddelde lingte - Polymerase | 67.995 | 89.379 | 49.664 | 66.433 |
| Gemiddelde lingte-Subreads | 15.866 | 21.036 | 11.657 | 16.012 |
| Gemiddelde lingte-CCS | 14.489 | 19.074 | 8.575 | 14.655 |
Gegevens generearre út 16 CLR-sellen (fan 76 soarten)
| DATA-PacBio-CLR-30Kb | Trochsneed | Max | Min | Mediaan |
| Yield - 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 |
| Gemiddelde lingte - Polymerase | 22.063 | 48.886 | 8.747 | 21.555 |
| Gemiddelde lingte-Subreads | 17.720 | 27.225 | 8.293 | 17.779 |
2.Data QC - DemoStatistiken oer gegevens opbringst
| Foarbyld | ccs Lês Num | Totale ccs-bases (bp) | ccs Reads N50 (bp) | ccs gemiddelde lingte (bp) | ccs langste lêzen (bp) | subreads Bases (bp) | ccs rate (%) |
| PB_BMKxxx | 3.444.159 | 54.164.122.586 | 15.728 | 15.726 | 36.110 | 863.326.330.465 | 6.27 |
