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Oloa

Eukaryotic mRNA fa'asologa-Illumina

mRNA fa'asologa e mafai ai ona fa'ailoaina uma mRNA fa'aliliu mai sela i lalo o tulaga fa'apitoa.Ose tekonolosi malosi mo le fa'aaliaina o fa'amatalaga fa'aaliga o ga'o, fa'asologa o kenera ma faiga mole mole o nisi gaioiga fa'aola.E oʻo mai i le taimi nei, o le mRNA sequencing ua faʻaaogaina lautele i suʻesuʻega faʻavae, suʻesuʻega falemaʻi, atinaʻeina o vailaʻau, ma isi.

Fa'avae: Illumina NovaSeq 6000


Au'aunaga Fa'amatalaga

Bioinformatics

I'uga Fa'aaliga

Tulaga lelei

ØSili Aʻoaʻoga: E sili atu i le 200,000 faʻataʻitaʻiga ua faʻatautaia i le BMK e aofia ai ituaiga faʻataʻitaʻiga eseese, e aofia ai le aganuʻu cell, tissue, sua o le tino, ma isi ma sili atu i le 7,000 mRNA-Seq poloketi tapunia e aofia ai vaega eseese o suʻesuʻega.

ØFa'atonuga lelei faiga: O vaega autu e pulea lelei e ala i laasaga uma e aofia ai sauniuniga fa'ata'ita'iga, tapenaga o faletusi, fa'asologa ma bioinformatics o lo'o i lalo o le mata'ituina lelei ina ia mafai ona tu'uina atu i'uga maualuga.

ØTele fa'amaumauga o lo'o avanoa mo fa'amatalaga galuega ma su'esu'ega fa'atamaoaigaina e fa'ataunu'u ai sini su'esu'ega eseese.

ØAu'aunaga ina ua mae'a fa'atau: Au'aunaga pe a mae'a fa'atauga e aoga mo le 3 masina pe a mae'a galuega, e aofia ai le tulitatao o poloketi, fa'alavelave fa'alavelave, fa'ai'uga Q&A, ma isi.

Fa'ata'ita'iga Mana'oga ma le Tu'uina atu

Faletusi Fuafuaga fa'asologa Fa'amatalaga fautuaina Puleaina lelei
Poly A fa'atamaoaigaina Illumina PE150

6 Gb

Q30≥85%

Fa'ata'ita'iga Mana'oga:

Nucleotides:

Mama atoatoa Amiotonu Aofaiga
OD260/280≥1.7-2.5 OD260/230≥0.5-2.5Fa'atapula'a pe leai se polotini po'o le DNA fa'aleagaina o lo'o fa'aalia ile gel. Mo la'au: RIN≥6.5;Mo manu: RIN≥7;28S/18S≥1.0;fa'atapula'a pe leai se maualuga maualuga Conc.≥30 ng/μl;Volum ≥ 10 μl;Aofa'i ≥ 1.5 μg

Tisu: Mamafa(mamago):≥1 g
*Mo mea e la'ititi ifo i le 5 mg, matou te fautuaina e lafo le moli fa'aaisa (i le vai nitrogen) fa'ata'ita'iga o mea'ai.

Taofi le sela:Faitauga sela = 3×106- 1×107
* Matou te fautuaina e lafo le lysate sela fa'aaisa.Afai e la'ititi ifo le numera o le sela nai lo le 5×105, e fautuaina le fa'a'aisa fa'aisa i le vai, lea e sili ona lelei mo le fa'a'aisa.

Su'ega toto:Volume≥1 ml

Microorganism:Massa ≥ 1 g

Fautuaina Fa'ata'ita'iga Tu'uina atu

Containe: 2 ml centrifuge tube (E le fautuaina le pepa apa)

Fa'ailoga fa'ata'ita'iga: Vaega+fa'atusa eg A1, A2, A3;B1, B2, B3... ...

utaina:

  1. Fa'amago-aisa: E mana'omia ona teu fa'ata'ita'i i taga ma tanu i le aisa mago.
  2. RNAstable tubes: RNA samples e mafai ona fa'amago ile RNA stabilization tube(fa'ata'ita'iga RNAstable®) ma lafo ile potu vevela.

Auala o Galuega Au'aunaga

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Fa'ata'ita'iga mamanu

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Fa'ata'ita'iga tu'uina atu

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RNA su'ega

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Fau faletusi

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Fa'asologa

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Iloiloga fa'amaumauga

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Au'aunaga pe a uma ona fa'atau


  • Muamua:
  • Sosoo ai:

  • Bioinformatics

    2(1)

    Eukaryotic mRNA su'esu'ega fa'asologa o galuega

    Bioinformatics

    ØPulea lelei fa'amatalaga mata'utia

    ØFa'asinoga genome alignment

    ØSu'esu'ega fa'asologa o tusitusiga

    ØFa'amatalaga fa'atusa

    ØSu'esu'ega fa'amatalaga eseese

    ØFa'amatalaga galuega ma fa'atamaoaigaina

    1.mRNA Fa'amatalaga Saturation curve

    3(1)

    2.Su'esu'ega fa'aaliga eseese-Volcano plot

    4(1)

    3.KEGG fa'amatalaga ile DEGs

    5(1)

    4.GO fa'avasegaga ile DEGs

    6(1)

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