DNA/RNA fa'asologa -PacBio Sequencer
Vaega
E lua faiga fa'asologa i luga ole fa'asologa o le PacBio: Fa'aauau le Faitau umi (CLR) ma le Faitau Fa'atasiga Fa'ata'amilo (CCS)
| Fa'asologa Fa'asologa | Tele o Faletusi | Fa'amatalaga Fa'amatalagaTu'u (Per Sel) | Fa'avae tasiSa'o | Talosaga |
| CLR | 20Kb, 30Kb, ma isi. | 80 Gb i le 130 Gb | Pe tusa.85% | De novo, valaau SV, ma isi. |
| CCS | 15-20 Kb | 14 i le 40 Gb/Cell (Sequel II) 70 i le 110 Gb/Cell (Revio) Fa'alagolago i fa'ata'ita'iga | Pe tusa.99% | De novo, SNP/Indel/SV vala'au, Iso-Seq, |
Fa'atusatusaga o fa'atinoga ma foliga o Revio ma le Fa'asologa II
| Tulaga | Fa'asologa II faiga | Revio faiga | Fa'ateleina |
| Maualuga maualuga | 8 miliona ZMWs | 25 miliona ZMWs | 3x |
| Tuto'atasi tulaga | 1 | 4 | 4x |
| Puupuu taimi tamo'e | 30 itula | 24 itula | 1.25x |
| 30X HiFi genome tagata / tausaga | 88 | 1,300 | 15x lautele |
Tulaga Auaunaga
● I luga ole 8 tausaga le poto masani ile PacBio sequencing platform ma le faitau afe o galuega tapunia ma ituaiga eseese.
● Faʻapipiʻiina atoatoa i faʻasologa fou o PacBio Sequencing, Revio e faʻamautinoa ai le lava o le faʻasologa o le gaosiga.
● Faʻavave le taimi liliu, maualuga le maua o faʻamaumauga ma sili atu faʻamatalaga saʻo.
● Fa'asoa i le fiaselau o fa'asalalauga fa'atatau i le PacBio.
Fa'ata'ita'iga Manaoga
| Ituaiga Fa'ata'ita'i | Aofaiga | Fa'atotonuga （Qubit®） | Volume | mama | O isi |
| Genomic DNA | Fa'alagolago i Fa'amatalaga Manaomia | ≥50 ng/μl | ≥15μl | OD260/280=1.7-2.2; OD260/230=1.8-2.5; manino tumutumu ile 260 nm，leai ni mea leaga | E manaʻomia ona fua le faʻatonuga e Qubit ma Qubit / Nanopore = 0.8-2.5 |
| RNA atoa | ≥1.2μg | ≥120 ng/μl | ≥15μl | OD260/280=1.7-2.5; OD260/230=0.5-2.5;leai ni mea leaga | RIN tau ≥7.5 5≥28S/18S≥1 |
Au'aunaga Galuega
Fa'ata'ita'iga sauniuniga
Fausia faletusi
Fa'asologa
Iloiloga fa'amaumauga
Tuuina atu o poloketi
1.In-fale Fa'amatalaga Tuuina atu
Fa'amaumauga na maua mai sela CCS 63 (mai le 26 ituaiga)
| FA'AMATALAGA-PacBio-CCS-15 Kb | averesi | Max | Min | Median |
| Tu'u - su'esu'e (Gb) | 421.12 | 544.27 | 221.38 | 426.58 |
| Maua - CCS(Gb) | 25.93 | 38.59 | 10.86 | 25.43 |
| Polymerase N50 | 145,651 | 175,430 | 118,118 | 144,689 |
| Laiti faitau N50 | 17,509 | 23,924 | 12,485 | 17,584 |
| CCS N50 | 14,490 | 19,034 | 9,876 | 14,747 |
| Ole umi ole umi-Polymerase | 67,995 | 89,379 | 49,664 | 66,433 |
| Aveta'i umi-Subreads | 15,866 | 21,036 | 11,657 | 16,012 |
| Uiga masani-CCS | 14,489 | 19,074 | 8,575 | 14,655 |
Fa'amaumauga na maua mai sela CLR 16 (mai le 76 ituaiga)
| DATA-PacBio-CLR-30Kb | averesi | Max | Min | Median |
| Tu'u - su'esu'e (Gb) | 142.20 | 291.40 | 50.55 | 142.49 |
| Polymerase N50 | 39,456 | 121,191 | 15,389 | 35,231 |
| Laiti faitau N50 | 28,490 | 41,012 | 14,430 | 29,063 |
| Ole umi ole umi-Polymerase | 22,063 | 48,886 | 8,747 | 21,555 |
| Aveta'i umi-Subreads | 17,720 | 27,225 | 8,293 | 17,779 |
2. Fa'amatalaga QC - Fa'aaligaFuainumera i luga o fa'amaumauga maua
| Fa'ata'ita'iga | ccs Faitau Nu | Aofa'i ccs Fa'avae (bp) | ccs Faitau N50 (bp) | ccs Uiga Umi (bp) | ccs Faitau Sili (bp) | subreads Fa'avae (bp) | fua faatatau ccs(%) |
| PB_BMKxxx | 3,444,159 | 54,164,122,586 | 15,728 | 15,726 | 36,110 | 863,326,330,465 | 6.27 |
