DNA/RNA sequentia -PacBio Sequencer
Features
Duo sequentes modos in PacBio sequencer: Continuus Long Read (CLR) et Consensus Circularis Read (CCS)
| Sequencing Modus | Library Size | Theoretical DataCede (Per Cell) | Unius basisSagacitas | Applications |
| CLR | 20Kb, 30Kb, etc. | 80 Gb ad 130 Gb | Proxime.85% | De novo, SV vocatum , etc. |
| CCS | 15-20 Kb | 14 ad 40 Gb/Cell (Sequel II) 70 ad 110 Mb/Cell (Revio) Exempla dependet | Proxime.99% | De novo, SNP/Indel/SV vocatio, Iso-Seq; |
Comparatio executionis et lineamenta Revio et Sequel II
| Terms | Sequentia II systema | Revio systema | Augere |
| Superiore densitate | VIII decies ZMWs | 25 decies centena millia ZMWs | 3x |
| Gradus independentes | 1 | 4 | 4x |
| Breviori currere temporibus | XXX horas | XXIV horas | 1.25x |
| 30X HiFi genomes human/annus | 88 | 1,300 | 15x altiore |
Servitium Commoda
● Plus 8 annorum experientia in PacBio sequencing suggestum cum millibus inceptis clausis cum variis speciebus.
● Exstructus cum recentissimis suggestis PacBio Sequencing, Revio spondere sat sequendo throughput.
● Velocius vertere circa tempus, superior notitia cedat et accuratior notitia.
● Contributed in centenis impulsus PacBio-substructio in publicationibus.
Sample Requisita
| Sample Type | Moles | Retrahitur Qubit®） | Magnitudo | Puritas | alii |
| Genomic DNA | Pendent Data opus est | ≥50 ng/μl | ≥15μl | OD260/280=1.7-2.2; OD260/230=1.8-2.5； Patet apicem at 260 nm，nulla contagione | Intentio metiri debet ab Qubit et 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；nulla contagione | RIN valor ≥7.5 5≥28S/18S≥1 |
Service Workflow
Sample praeparatio
Library construction
Sequencing
Analysis
Project traditio
1.In domo data cede
Data ex cellulis 63 CCS generatis (ex 26 speciebus)
| DATA-PacBio-CCS-15 Kb | Mediocris | Max | Min | Medus |
| Cede - 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 |
| Mediocris longitudinis Polymerase | 67,995 | 89,379 | 49,664 | 66,433 |
| Mediocris longitudinis Subreads | 15,866 | 21,036 | 11,657 | 16,012 |
| Mediocris longitudinis CCS | 14,489 | 19,074 | 8,575 | 14,655 |
Data ex 16 CLR cellulis generata (e 76 speciebus)
| DATA-PacBio-CLR-30Kb | Mediocris | Max | Min | Medus |
| Cede - 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 |
| Mediocris longitudinis Polymerase | 22,063 | 48,886 | 8,747 | 21,555 |
| Mediocris longitudinis Subreads | 17,720 | 27,225 | 8,293 | 17,779 |
2.Data QC - DemoStatistics in notitia cede
| Sample | CCS legit Num | Summa CCS Bases sitæ (bp) | CCS legit N50 (bp) | CCS Mean Longitudo (bp) | CCS maximus Read (bp) | subreads Bases sitæ (bp) | CCS rate (%) |
| PB_BMKxxx | 3,444,159 | 54,164,122,586 | 15,728 | 15,726 | 36,110 | 863,326,330,465 | 6.27 |
