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Cov khoom

Cov tsev qiv ntawv Illumina uas tau ua tiav lawm

Cov thev naus laus zis Illumina sequencing, raws li Sequencing by Synthesis (SBS), yog ib qho kev tsim kho tshiab NGS thoob ntiaj teb, uas yog lub luag haujlwm rau kev tsim ntau dua 90% ntawm cov ntaub ntawv sequencing thoob ntiaj teb. Lub hauv paus ntsiab lus ntawm SBS suav nrog kev thaij duab fluorescently labeled reversible terminators thaum txhua dNTP raug ntxiv, thiab tom qab ntawd cleaved kom tso cai rau kev koom ua ke ntawm lub hauv paus tom ntej. Nrog rau tag nrho plaub reversible terminator-bound dNTPs uas muaj nyob rau hauv txhua lub voj voog sequencing, kev sib tw ntuj tsim txo qis kev sib xyaw ua ke. Cov thev naus laus zis ntau yam no txhawb nqa ob qho tib si nyeem ib leeg thiab khub-kawg tsev qiv ntawv, ua kom haum rau ntau yam kev siv genomic. Illumina sequencing lub peev xwm siab-throughput thiab qhov tseeb tso nws ua lub hauv paus tseem ceeb hauv kev tshawb fawb genomics, txhawb nqa cov kws tshawb fawb kom daws qhov nyuaj ntawm genomes nrog cov ntsiab lus tsis sib xws thiab kev ua haujlwm zoo.

Peb qhov kev pabcuam sequencing uas tau ua tiav lawm ua rau cov neeg siv khoom tuaj yeem npaj cov tsev qiv ntawv sequencing los ntawm ntau qhov chaw (mRNA, tag nrho cov genome, amplicon, 10x cov tsev qiv ntawv, thiab lwm yam). Tom qab ntawd, cov tsev qiv ntawv no tuaj yeem xa mus rau peb lub chaw sequencing rau kev tswj xyuas zoo thiab sequencing hauv Illumina platforms.


Cov Lus Qhia Txog Kev Pabcuam

Cov txiaj ntsig ntawm kev sim

Cov yam ntxwv

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

 

 

Cov Kev Pabcuam Zoo

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.

 

 

Cov Qauv Platforms

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

Cov Qauv Yuav Tsum Tau Ua

 

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.

Qhov Loj ntawm Tsev Qiv Ntawv (Daim Ntawv Qhia Qhov Siab Tshaj Plaws)

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.

Thov hu rau peb yog tias koj cov qauv tsis ua tau raws li cov khoom siv pib.

Kev Ua Haujlwm Pabcuam

kev npaj qauv

Kev tswj xyuas zoo ntawm tsev qiv ntawv

Kev txheeb xyuas kab ke

Kev txheeb xyuas kab ke

Kev tshuaj xyuas cov ntaub ntawv

Kev tswj xyuas cov ntaub ntawv zoo

Qauv QC

Kev xa khoom ntawm qhov project


  • Yav dhau los:
  • Tom ntej no:

  • Daim ntawv qhia txog QC ntawm tsev qiv ntawv

    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.

     

    Daim ntawv qhia txog kev txheeb xyuas QC

     

    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

    A9

    Daim Duab 2. Kev faib cov ntsiab lus tseem ceeb

    A10

    Daim Duab 3. Kev faib tawm ntawm cov ntsiab lus nyeem hauv cov ntaub ntawv sequencing

    A11

     

    tau ib daim ntawv hais nqe

    Sau koj cov lus ntawm no thiab xa tuaj rau peb

    Xa koj cov lus rau peb: