Eukaryotic mRNA Sequencing-Illumina

mRNA fa'asologa e mafai ai ona fa'ailoaina uma mRNA fa'aliliu mai sela i lalo o tulaga fa'apitoa.Ose fa'atekonolosi mamana mo le fa'aaliaina o fa'amatalaga fa'amatalaga o le kenera, 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.

Tulaga: Illumina NovaSeq Platform

Faafesootai matou

Au'aunaga Fa'amatalaga

Bioinformatics

I'uga Fa'aaliga

Tulaga lelei

● Aʻoaʻoga Maualuga: 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, vai o le tino, ma isi mea ma sili atu i le 7,000 mRNA-Seq poloketi tapunia e aofia ai vaega eseese o suʻesuʻega.

● Fa'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.

● Tele 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.

● Au'aunaga ina ua mae'a fa'atau: Au'aunaga ina ua 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:

Conc.(ng/μl)

Aofa'i (μg)

Mama

Amiotonu

≥ 20

≥ 0.5

OD260/280=1.7-2.5

OD260/230=0.5-2.5

Fa'atapula'a pe leai se polotini po'o le fa'aleagaina o DNA o lo'o fa'aalia ile gel.

Mo laau toto: RIN≥6.5;

Mo manu: RIN≥7.0;

5.0≥28S/18S≥1.0;

fa'atapula'a pe leai se maualuga maualuga

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

Taofi le sela:Faitauga sela = 3×10⁶- 1×10⁷
* Matou te fautua atu e lafo le lysate cell frozen.I le tulaga o lena sela e laʻititi ifo nai lo le 5x105.

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:

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