Usoro DNA/RNA -PacBio Sequencer
Atụmatụ
Ụdị usoro usoro abụọ na usoro PacBio: Ọgụgụ ogologo na-aga n'ihu (CLR) na agụ nkwenye okirikiri (CCS)
| Usoro usoro | Nha ọba akwụkwọ | Data usoro iwuMpụta (kwa cell) | Otu-baseizi ezi | Ngwa |
| CLR | 20Kb, 30kb, wdg. | 80GB ruo 130GB | Ihe ruru.85% | De novo, SV na-akpọ, wdg. |
| CCS | 15-20 Kb | 14 ruo 40 Gb/cell (Sequel II) 70 ruo 110 Gb/cell (Revio) Na-adabere na samples | Ihe ruru.99% | De novo, SNP/Indel/SV na-akpọ, Iso-Seq, |
Ntụle nke arụmọrụ na atụmatụ nke Revio na Sequel II
| Usoro | Usoro nke usoro II | Sistemụ Revio | Mmụba |
| Njupụta dị elu | 8 nde ZMW | 25 nde ZMW | 3x |
| Usoro nke onwe | 1 | 4 | 4x |
| Oge ịgba ọsọ dị mkpụmkpụ | awa 30 | awa 24 | 1.25x |
| 30X HiFi genome mmadụ / afọ | 88 | 1,300 | 15x n'ozuzu |
Uru ọrụ
● Ihe karịrị 8 afọ ahụmahụ na PacBio sequencing n'elu ikpo okwu na ọtụtụ puku mechiri emechi oru ngo na dị iche iche ụdị.
● Ejiri usoro PacBio Sequencing kachasị ọhụrụ, Revio na-ekwe nkwa mmepụta usoro zuru oke.
● Oge ntụgharị ọsọ ọsọ, mkpụrụ data dị elu yana data ziri ezi.
● Enyere aka n'ọtụtụ narị akwụkwọ ndị dabeere na PacBio nwere mmetụta dị elu.
Ihe atụ chọrọ
| Ụdị Nlereanya | Ọnụ ego | Ntinye uche (Qubit®) | Olu | Ịdị ọcha | Ndị ọzọ |
| Genomic DNA | Dabere na data chọrọ | ≥50 ng/μl | ≥15 ml | OD260/280=1.7-2.2; OD260/230=1.8-2.5; Kpochapụ elu na 260 nmenweghị mmetọ | Ekwesịrị iji Qubit na Qubit/Nanopore tụọ ntinye uche = 0.8-2.5 |
| Mkpokọta RNA | ≥1.2μg | ≥120 ng/μl | ≥15 ml | OD260/280=1.7-2.5; OD260/230=0.5-2.5;enweghị mmetọ | Ọnụ ego RIN ≥7.5 5≥28S/18S≥1 |
Usoro ọrụ ọrụ
Nkwadebe ihe atụ
Ịrụ ụlọ akwụkwọ
Usoro
Nyocha data
Nnyefe oru ngo
1.N'ụlọ Data Mpụta
Data sitere na sel 63 CCS (site na ụdị 26)
| DATA-PacBio-CCS-15 Kb | Nkezi | Oke | Min | Ọkara |
| Mpụta - agụpụta (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 |
| Ọ na-agụ N50 | 17,509 | 23,924 | 12,485 | 17,584 |
| CCS N50 | 14,490 | 19,034 | 9,876 | 14,747 |
| Nkezi ogologo-Polymerase | 67,995 | 89,379 | 49,664 | 66,433 |
| Nkezi ogologo-Achịcha ọka | 15,866 | 21,036 | 11,657 | 16,012 |
| Nkezi ogologo-CCS | 14,489 | 19,074 | 8,575 | 14,655 |
Data sitere na mkpụrụ ndụ 16 CLR (site na ụdị 76)
| DATA-PacBio-CLR-30Kb | Nkezi | Oke | Min | Ọkara |
| Mpụta - agụpụta (Gb) | 142.20 | 291.40 | 50.55 | 142.49 |
| Polymerase N50 | 39,456 | 121,191 | 15,389 | 35,231 |
| Ọ na-agụ N50 | 28,490 | 41,012 | 14,430 | 29,063 |
| Nkezi ogologo-Polymerase | 22,063 | 48,886 | 8,747 | 21,555 |
| Nkezi ogologo-Achịcha ọka | 17,720 | 27,225 | 8,293 | 17,779 |
2.Data QC - ngosiStatistics na mkpụrụ data
| Ihe atụ | ccs na-agụ Ọgụ | Mgbakọta ccs ntọala (bp) | ccs na-agụ N50 (bp) | ccs Ogologo Ogologo (bp) | ccs Kariri ọgụgụ (bp) | ntọala subreads (bp) | Ọnụego ccs (%) |
| PB_BMKxxx | 3,444,159 | 54,164,122,586 | 15,728 | 15,726 | 36,110 | 863,326,330,465 | 6.27 |
