Te raupapa DNA/RNA -PacBio Sequencer
Ngā āhuatanga
E rua nga aratau raupapa i runga i te PacBio raupapa: Panui Roa Tonu (CLR) me te Panui Whakaaetanga Porohita (CCS)
| Aratau Raupapa | Rahi Whare Pukapuka | Raraunga TikangaHua (Ia Pūtau) | Turanga-tahiTika | Nga tono |
| CLR | 20Kb, 30Kb, aha atu. | 80 Gb ki te 130 Gb | Approx.85% | De novo, waea SV, etc. |
| CCS | 15-20 Kb | 14 ki te 40 Gb/Pūtau (Sequel II) 70 ki te 110 Gb/Putau (Revio) Kei runga i nga tauira | Approx.99% | De novo, Waea SNP/Indel/SV, Iso-Seq, |
Te whakataurite o nga mahi me nga ahuatanga o Revio me Sequel II
| Nga tikanga | Pūnaha Paarua II | Pūnaha Revo | Whakanuia |
| Kiato teitei ake | 8 miriona ZMW | 25 miriona ZMW | 3x |
| Nga wahanga motuhake | 1 | 4 | 4x |
| He wa poto ake te oma | 30 haora | 24 haora | 1.25x |
| 30X HiFi ira tangata / tau | 88 | 1,300 | 15x katoa |
Painga Ratonga
● Neke atu i te 8 tau te wheako i runga i te papa raupapa raupapa PacBio me nga mano tini kaupapa kati me nga momo momo.
● Kua rite rawa ki nga papaaho PacBio Sequencing hou, Revio ki te whakapumau i te tika o te whakaputanga raupapa.
● He tere ake te hurihanga, he nui ake te tukunga o nga raraunga me nga raraunga tika ake.
● I koha i roto i nga rau o nga whakaputanga-nui-a-PacBio.
Nga Whakaritenga Tauira
| Momo Tauira | Te nui | Kukū （Qubit®） | Rōrahi | Maamaa | Ko etahi atu |
| DNA genomic | Kei runga i nga whakaritenga Raraunga | ≥50 ng/μl | ≥15μl | OD260/280=1.7-2.2; OD260/230=1.8-2.5； Maama te tihi i te 260 nm，karekau he poke | Me ine te kukū ma te Qubit me te Qubit/Nanopore = 0.8-2.5 |
| RNA katoa | ≥1.2μg | ≥120 ng/μl | ≥15μl | OD260/280=1.7-2.5; OD260/230=0.5-2.5；karekau he poke | Uara RIN ≥7.5 5≥28S/18S≥1 |
Rerengamahi Ratonga
Te whakarite tauira
Te hanga whare pukapuka
Whakaraupapa
Te tātari raraunga
Te tuku kaupapa
1.I roto-whare Tukunga Raraunga
Raraunga i hangaia mai i nga pūtau CCS 63 (mai i nga momo 26)
| RARAUNGA-PacBio-CCS-15 Kb | Wawaenga | Max | Min | Takawaenga |
| Hua - panui-iti (Gb) | 421.12 | 544.27 | 221.38 | 426.58 |
| Tukua - CCS(Gb) | 25.93 | 38.59 | 10.86 | 25.43 |
| Polymerase N50 | 145,651 | 175,430 | 118,118 | 144,689 |
| Nga panui iti N50 | 17,509 | 23,924 | 12,485 | 17,584 |
| CCS N50 | 14,490 | 19,034 | 9,876 | 14,747 |
| Roa Toharite-Polymerase | 67,995 | 89,379 | 49,664 | 66,433 |
| Roa Toharite-Paaroto | 15,866 | 21,036 | 11,657 | 16,012 |
| Roa Toharite-CCS | 14,489 | 19,074 | 8,575 | 14,655 |
Raraunga i hangaia mai i nga pūtau CLR 16 (mai i nga momo 76)
| DATA-PacBio-CLR-30Kb | Wawaenga | Max | Min | Takawaenga |
| Hua - panui-iti (Gb) | 142.20 | 291.40 | 50.55 | 142.49 |
| Polymerase N50 | 39,456 | 121,191 | 15,389 | 35,231 |
| Nga panui iti N50 | 28,490 | 41,012 | 14,430 | 29,063 |
| Roa Toharite-Polymerase | 22,063 | 48,886 | 8,747 | 21,555 |
| Roa Toharite-Paaroto | 17,720 | 27,225 | 8,293 | 17,779 |
2.QC Raraunga – WhakaaturangaNga tatauranga mo te tukunga raraunga
| Tauira | ccs Panui Num | Tapeke ccs turanga (bp) | ccs Panui N50 (bp) | ccs Roa Tikanga (bp) | ccs Pānui Roa Rawa (bp) | Panui iti (bp) | reeti ccs(%) |
| PB_BMKxxx | 3,444,159 | 54,164,122,586 | 15,728 | 15,726 | 36,110 | 863,326,330,465 | 6.27 |
