●Cov platforms:Illumina NovaSeq 6000 thiab NovaSeq X Plus
●Cov hom kev txheeb xyuas:PE150 thiab PE250
●Kev tswj xyuas zoo ntawm cov tsev qiv ntawv ua ntej kev txheeb xyuas cov ntaub ntawv
●Kev tshuaj xyuas cov ntaub ntawv sequencing QC thiab kev xa khoom:Kev xa daim ntawv qhia QC thiab cov ntaub ntawv raw hauv hom ntawv fastq tom qab demultiplexing thiab lim Q30 nyeem
●Kev ua tau ntau yam ntawm cov kev pabcuam Sequencing:tus neeg siv khoom xaiv tau los ua qhov kev txiav txim siab raws li txoj kab, cov flow cell, lossis raws li qhov ntau ntawm cov ntaub ntawv xav tau (ib nrab txoj kab sequencing).
●Muaj kev paub ntau yam ntawm Illumina sequencing platform:nrog ntau txhiab qhov project kaw nrog ntau hom tsiaj.
●Kev xa daim ntawv qhia txog kev txheeb xyuas QC:nrog rau cov ntsuas zoo, qhov tseeb ntawm cov ntaub ntawv thiab kev ua tau zoo tag nrho ntawm qhov project sequencing.
●Cov txheej txheem txheeb xyuas cov neeg laus:nrog lub sijhawm luv luv tig rov qab.
●Kev Tswj Xyuas Zoo: peb siv cov kev cai QC nruj heev kom lav qhov xa khoom zoo tas li.
| Platform | Flow Cell | Hom kev txheeb xyuas | Chav tsev | Kwv yees cov zis tawm |
| NovaSeq X | 10B (8 txoj kab) | PE150 | Txoj Kev Ib Leeg Txoj Kev Ib Nrab | 375Gb / Txoj Kev |
| 25B (8 txoj kab) | PE150 | Txoj Kev Ib Leeg Txoj Kev Ib Nrab | 1000 Gb/Txoj Kab | |
| NovaSeq 6000 | SP Flow cell (2 txoj kab) | PE250 | Flow Cell Txoj Kev Ib Leeg Txoj Kev Ib Nrab | 325-400 M nyeem / Txoj kab |
| S4 Flow cell (4 txoj kab) | PE150 | Flow Cell Txoj Kev Ib Leeg Txoj Kev Ib Nrab | ~800 Gb / Txoj Kab |
| Tus Nqi Ntaub Ntawv (Gb) | Conc. (qPCR) PE150 | Conc. (qPCR) PE250 | Ntim | |
| Txoj Kev Ib Nrab
| G ≤ 10 | ≥ 1 nM | ≥ 2nM | ≥ 25 μl |
| 10 < G ≤ 50 | ≥ 2 nM | ≥ 3nM | ≥ 25 μl | |
| 50 < G ≤ 100 | ≥ 3 nM | ≥ 4nM | ≥ 25 μl | |
| G > 100 | ≥ 4 nM | ≥ 4nM | ≥ 25 μl | |
| Txoj Kev Ib Leeg | Ib Txoj Kab | ≥ 1.5 nM/Lub pas dej hauv tsev qiv ntawv | ≥ 2 nM/Lub pas dej hauv tsev qiv ntawv | ≥ 25 µL/pas dej hauv tsev qiv ntawv |
Ntxiv rau qhov concentration thiab tag nrho cov nyiaj, kuj xav tau tus qauv ncov tsim nyog.
Lus Cim: Kev txheeb xyuas txoj kab ntawm cov tsev qiv ntawv tsis muaj ntau haiv neeg xav tau PhiX spike-in kom ntseeg tau tias muaj kev hu xov tooj ruaj khov.
Peb xav kom xa cov tsev qiv ntawv uas tau muab ua ke ua qauv rau kev txheeb xyuas txoj kab. Yog tias koj xav tau BMKGENE los ua kev sib sau ua ke ntawm tsev qiv ntawv, thov saib cov kev cai ntawm tsev qiv ntawv rau kev txheeb xyuas ib nrab txoj kab.
Rau PE150 sequencing, qhov loj ntawm cov ntxig yuav tsum yog 300–400 bp (tsis suav nrog cov adapters); rau PE250 sequencing, qhov loj ntawm cov ntxig yuav tsum yog 150–500 bp (tsis suav nrog cov adapters).
Cov tsev qiv ntawv yuav tsum muaj ib lub ncov tseem ceeb, tsis muaj kev sib kis ntawm lub adapter thiab tsis muaj cov primer dimers.
Muaj ib daim ntawv qhia txog qhov zoo ntawm lub tsev qiv ntawv ua ntej kev txheeb xyuas, kev ntsuas tus nqi ntawm lub tsev qiv ntawv, thiab kev faib ua feem.
Rooj 1. Cov ntaub ntawv txheeb cais ntawm cov ntaub ntawv sequencing.
| Tus qauv ID | BMKID | Cov ntawv nyeem tsis tau ua tiav | Cov Ntaub Ntawv Raw (bp) | Nyeem huv si (%) | Q20 (%) | Q30 (%) | GC (%) |
| C_01 | BMK_01 | 22,870,120 | 6,861,036,000 | 96.48 | 99.14 | 94.85 | 36.67 |
| C_02 | BMK_02 | 14,717,867 | 4,415,360,100 | 96.00 | 98.95 | 93.89 | 37.08 |
Daim Duab 1. Kev faib tawm zoo raws li kev nyeem hauv txhua tus qauv
Daim Duab 2. Kev faib cov ntsiab lus tseem ceeb
Daim Duab 3. Kev faib tawm ntawm cov ntsiab lus nyeem hauv cov ntaub ntawv sequencing