Mode Sequencing | Ukuran Perpustakaan | Data TéoritisNgahasilkeun (Per Sél) | Dasar tunggalAkurasi | Aplikasi |
CLR | 20 Kb, 30 Kb, jsb. | 80 Gb ka 130 Gb | Kira-kira.85% | De novo, SV nelepon, jsb. |
CCS | 15-20 Kb | 14 nepi ka 40 Gb/Sél (Sequel II) 70 nepi ka 110 Gb/Sél (Revio) Gumantung kana sampel | Kira-kira.99% | De novo, Telepon SNP/Indel/SV, Iso-Seq, |
Sarat | Sistem Sekuel II | Sistim Revo | Naékkeun |
Kapadetan luhur | 8 juta ZMWs | 25 juta ZMWs | 3x |
Tahap mandiri | 1 | 4 | 4x |
waktos ngajalankeun pondok | 30 jam | 24 jam | 1,25x |
30X HiFi génom manusa / taun | 88 | 1.300 | 15x sakabéh |
● Leuwih 8 taun pangalaman dina platform sequencing PacBio kalawan rébuan proyék katutup jeung sagala rupa spésiés.
● Pinuh dilengkepan platform PacBio Sequencing panganyarna, Revio pikeun ngajamin throughput sequencing cukup.
● Waktos péngkolan langkung gancang, ngahasilkeun data anu langkung luhur sareng data anu langkung akurat.
● Kontribusi dina ratusan publikasi dumasar-PacBio dampak luhur.
Tipe Sampel | Jumlah | Konsentrasi (Qubit®) | Jilid | Kasucian | Lain |
DNA génomik | Gumantung kana Sarat Data | ≥50 ng/μl | ≥15μl | OD260/280 = 1.7-2.2; OD260/230 = 1.8-2.5; Puncak jelas dina 260 nm,euweuh contaminations | Konsentrasi kedah diukur ku Qubit sareng Qubit/Nanopore = 0.8-2.5 |
RNA total | ≥1,2 μg | ≥120 ng/μl | ≥15μl | OD260/280 = 1.7-2.5; OD260/230 = 0,5-2,5;euweuh contaminations | nilai RIN ≥7.5 5≥28S/18S≥1 |
1.In-imah Data ngahasilkeun
Data dihasilkeun tina 63 sél CCS (tina 26 spésiés)
DATA-PacBio-CCS-15 Kb | Rata-rata | Max | Min | Median |
Ngahasilkeun - subreads (Gb) | 421.12 | 544.27 | 221.38 | 426.58 |
Ngahasilkeun - CCS(Gb) | 25.93 | 38.59 | 10.86 | 25.43 |
Polimérase 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 |
Rata-rata panjangna-Polimérase | 67.995 | 89.379 | 49.664 | 66.433 |
Rata panjang-Subreads | 15.866 | 21.036 | 11.657 | 16.012 |
Rata-rata panjangna-CCS | 14.489 | 19.074 | 8.575 | 14.655 |
Data dihasilkeun tina 16 sél CLR (tina 76 spésiés)
DATA-PacBio-CLR-30Kb | Rata-rata | Max | Min | Median |
Ngahasilkeun - subreads (Gb) | 142.20 | 291.40 | 50.55 | 142.49 |
Polimérase N50 | 39.456 | 121.191 | 15.389 | 35.231 |
Subreads N50 | 28.490 | 41.012 | 14.430 | 29.063 |
Rata-rata panjangna-Polimérase | 22.063 | 48.886 | 8.747 | 21.555 |
Rata panjang-Subreads | 17.720 | 27.225 | 8.293 | 17.779 |
2. Data QC - DemoStatistik dina ngahasilkeun data
Sampel | ccs Bacaan Num | Total ccs Base (bp) | ccs Bacaan N50 (bp) | ccs Rata-rata Panjang (bp) | ccs Bacaan Pangpanjangna (bp) | Subbacaan Dasar (bp) | laju ccs(%) |
PB_BMKxxx | 3.444.159 | 54.164.122.586 | 15.728 | 15.726 | 36.110 | 863.326.330.465 | 6.27 |