条形 ibhanile-03

Iimveliso

[Ikopi] I-Eukaryotic mRNA Sequencing-NGS

Ukulandelelana kwe-mRNA, iteknoloji eguquguqukayo, inika amandla ukubhalwa ngokubanzi kwazo zonke ii-mRNA transcripts ngaphakathi kweeseli phantsi kweemeko ezithile. Ngezicelo zayo ezibanzi, esi sixhobo siphambili sityhila iiprofayili zokubonakaliswa kwezakhi zofuzo ezintsonkothileyo, izakhiwo zezakhi zofuzo, kunye neendlela zemolekyuli ezinxulumene neenkqubo ezahlukeneyo zebhayoloji. Isetyenziswa kakhulu kuphando olusisiseko, ukuxilongwa kwezonyango, kunye nophuhliso lwamayeza, ukulandelelana kwe-mRNA kunika ulwazi malunga nobunzima be-dynamics yeseli kunye nolawulo lwezakhi zofuzo, ivuselela umdla malunga namandla ayo kwiindawo ezahlukeneyo.

Iqonga: Illumina NovaSeq X; DNBSEQ-T7


Iinkcukacha zeNkonzo

I-Bioinformatics

Iziphumo zeDemo

Iimpapasho Eziphambili

Iimbonakalo

● Ukubanjwa kwe-poly mRNA ngaphambi kokulungiswa kwethala leencwadi

● Ukulungiselela ilayibrari ye-mRNA enokukhokela ukuze ukwazi ukufumana idatha yokulandelelana kwemigca ethile

● Uhlalutyo lwe-Bioinformatic lwendlela yokubonakaliswa kwezakhi zofuzo kunye nesakhiwo se-transcript

 

Iingenelo

●Ubuchule ObubanziIqela lethu licubungule iisampuli ezingaphezu kwama-600,000 kwi-BMKgene, kuquka iintlobo ezahlukeneyo zeesampuli ezifana neenkcubeko zeeseli, izicubu zomzimba, kunye nolwelo lomzimba. Sizisa amava amaninzi kuyo yonke iprojekthi kwaye sigqibe ngempumelelo iiprojekthi ze-mRNA-Seq ezingaphezu kwe-100,000 kwiindawo ezahlukeneyo zophando.

●Ulawulo Oluqinileyo Lomgangatho: Sisebenzisa amanqaku olawulo oluphambili kuzo zonke izigaba, ukusuka ekulungiseleleni isampuli kunye nethala leencwadi ukuya kulandelelwano kunye ne-bioinformatics. Olu hlolo olucokisekileyo luqinisekisa ukunikezelwa kweziphumo ezisemgangathweni ophezulu rhoqo, luqinisekisa ukuba iprojekthi yakho isezandleni ezithembekileyo.

●Ingcaciso ebanzi: Sisebenzisa iidathabheyisi ezininzi ukuze sichaze ngokusebenzayo iiGenes ezichazwe ngokuDifferentially Expressed Genes (DEGs) kwaye senze uhlalutyo lokufumisa oluhambelanayo. Le ndlela ibanzi inika ulwazi kwiinkqubo zeseli kunye neemolekyuli eziphantsi kwempendulo ye-transcriptome, ukuqinisekisa ukuba unolwazi olupheleleyo malunga neziphumo zeprojekthi yakho.

●Inkxaso Emva koThengiso: Ukuzibophelela kwethu kudlulela ngaphaya kokugqitywa kweprojekthi kunye nexesha leenkonzo emva kokuthengiswa kweenyanga ezi-3. Ngeli xesha, sinikezela ngokulandela iprojekthi, uncedo lokusombulula iingxaki, kunye neeseshoni ze-Q&A ukuphendula nayiphi na imibuzo enxulumene neziphumo.

Iimfuno zeSampuli kunye nokuHanjiswa

Ithala leeNcwadi Icebo lokulandelelanisa izinto Idatha iyacetyiswa Ulawulo lwemeko
I-Poly A etyebileyo

I-Illumina PE150

I-DNBSEQ-T7

6 -10 Gb

Q30≥85%

Iimfuno zeSampuli:

Iinukleotide:

Iimfuno zeSampuli eziNcinci:

 

I-Conc.(ng/μl)

Ubungakanani (μg)

Ubunyulu

Ingqibelelo

Ithala leeNcwadi eliQhelekileyo

≥ 10

≥ 0.2

OD260/280=1.7-2.5

OD260/230=0.5-2.5

Ungcoliseko lweproteni okanye lwe-DNA oluncinci okanye alubonakali kwijeli.

Kwizityalo: I-RIN≥4.0;

Kwizilwanyana: I-RIN≥4.5;

5.0≥28S/18S≥1.0;

ukuphakama okulinganiselweyo okanye okungekho siseko

Ithala leeNcwadi eliLawulayo

≥ 10

≥ 0.2

OD260/280=1.7-2.5

OD260/230=0.5-2.5

Ungcoliseko lweproteni okanye lwe-DNA oluncinci okanye alubonakali kwijeli.

Kwizityalo: I-RIN≥4.0;

Kwizilwanyana: I-RIN≥4.5;

5.0≥28S/18S≥1.0;

ukuphakama okulinganiselweyo okanye okungekho siseko

Izityalo Isilwanyana Ii-arthropod Igazi elipheleleyo Iiseli

Ingcambu, Isiqu okanye iPetal: 450 mg

Igqabi okanye Imbewu: 300 mg

Isiqhamo: 1.2 g

Intliziyo okanye Amathumbu: 300 mg

I-Viscera okanye ubuchopho: 240 mg

Izihlunu: 450 mg

Amathambo, Iinwele okanye Ulusu: 1g

Izinambuzane: 6g

I-Crustacea: 300 mg

Ityhubhu e-1 Iiseli ezili-106

Ukuhanjiswa kweSampuli okuCetyisiweyo

Isikhongozeli: Ityhubhu ye-centrifuge ye-2 ml (Akukhuthazwa ukusebenzisa i-tin foil)

Ilebheli yesampulu: Iqela + ikopi umz. A1, A2, A3; B1, B2, B3... ...

Ukuthunyelwa:

1. Umkhenkce owomileyo: Iisampulu kufuneka zipakishwe kwiingxowa zize zingcwatywe kumkhenkce owomileyo.

2. Iityhubhu zetafile ze-RNA: Iisampulu ze-RNA zingomiswa kwityhubhu yokuzinzisa i-RNA (umz. i-RNAstable®) kwaye zithunyelwe kubushushu begumbi.

Iimfuno zeSampuli kunye nokuHanjiswa

123v

Jonga Okungakumbi

q1
q2
q3

Ukuba unemibuzo malunga neemveliso zethu okanye uluhlu lwamaxabiso, nceda usishiyele i-imeyile yakho.
Siza kuqhagamshelana nathi kwiiyure ezingama-24.

 

  • Ngaphambili:
  • Okulandelayo:

  • I-Bioinformatics

    i-wps_doc_10

    I-Eukaryotic Umsebenzi wohlalutyo lokulandelelana kwe-mRNA

    I-Bioinformatics

    ØUlawulo lomgangatho wedatha eluhlaza

    ØUlungelelwaniso lwe-genome yesalathiso

    ØUhlalutyo lwesakhiwo semibhalo ebhaliweyo

    ØUbungakanani bokuchaza

    ØUhlalutyo lokubonakaliswa kokwahluka

    ØIsichazi-magama somsebenzi kunye nokuphucula

    Ulungelelwaniso lweeGenome ezibhekiselele kuzo

     

    图片1

     

    Ukwanda kwedatha

     

    3(1)

     

     

    Ulwalamano lwesampulu kunye novavanyo lweekopi zebhayoloji

     

    图片2

     

    IiGene ezichazwe ngokwahlukileyo (ii-DEG)

     

    4(1)

     

     

    Ingcaciso esebenzayo yee-DEG

     

    6(1)

     

     

     

    Ukuphuculwa kokusebenza kwe-DEGs

     

    umfanekiso we-4

     

    Hlola inkqubela phambili eququzelelwe ziinkonzo ze-BMKGene zokulandelelana kwe-eukaryotic NGS mRNA ngokusebenzisa ingqokelela yeempapasho ezikhethiweyo.

     

    UHuang, L. et al. (2023) 'I-Triclosan kunye ne-triclocarban zibuthathaka amandla okunuka e-goldfish ngokuthintela ukuqatshelwa kwevumba, ukuphazamisa ukudluliselwa kwesignali yokunuka, kunye nokuphazamisa ukucutshungulwa kolwazi lokunuka', uPhando lwaManzi, 233, iphe. 119736. doi: 10.1016/J.WATRES.2023.119736.

    UJia, LJ et al. (2023) 'I-Aspergillus fumigatus ithatha i-p11 yomntu ukuze iqondise ii-phagosomes eziqulathe isikhunta kwindlela engonakalisiyo', i-Cell Host kunye ne-Microbe, 31(3), iphe. 373-388.e10. doi: 10.1016/J.CHOM.2023.02.002.

    UJin, K. et al. (2022) 'Izinto zeTCP Transcription ezibandakanyekayo ekuphuhlisweni kweZihluma zeMa Bamboo (Dendrocalamus latiflorus Munro)', Frontiers in Plant Science, 13, p. 884443. doi: 10.3389/FPLS.2022.884443/BIBTEX.

    UWen, X. et al. (2022) 'I-genome ye-Chrysanthemum lavandulifolium kunye nendlela esebenza ngayo i-molecular engaphantsi kweentlobo ezahlukeneyo ze-capitulum', Uphando lwezoLimo, 9. doi: 10.1093/HR/UHAB022.

    UZhang, uYujie et al. (2023) 'I-cascade nanoreactor yokuphucula unyango lwe-sonodynamic kumhlaza we-colorectal ngokusebenzisa i-synergistic ROS augment kunye ne-autophagy blockage', Nano Today, 49, p. 101798. doi: 10.1016/J.NANTOD.2023.101798.

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