China Eukaryotic mRNA sequencing-Ụlọ ọrụ mmepụta ihe na ndị na-emepụta ihe |BMK

Eukaryotic mRNA usoro-Ilumina

Eukaryotic mRNA sequencing-Illumina Featured Image

usoro mRNA na-enyere aka ịkọwapụta mRNA niile edere site na sel n'okpuru ọnọdụ akọwapụtara.Ọ bụ teknụzụ siri ike maka ikpughe profaịlụ mkpụrụ ndụ ihe nketa, usoro mkpụrụ ndụ ihe nketa na usoro mkpụrụ ndụ nke ụfọdụ usoro ndu.Ruo ugbu a, usoro mRNA ejirila n'ọrụ n'ọtụtụ ebe na nyocha nyocha, nyocha ụlọ ọgwụ, mmepe ọgwụ, wdg.

Platform: Illumina NovaSeq 6000

Kpọtụrụ anyị

Nkọwa ọrụ

Bioinformatics

Nsonaazụ ngosi

Uru

ØAhụmahụ dị ukwuu: Ahazila ihe karịrị 200,000 sample na BMK na-ekpuchi ụdị ụdị dị iche iche, gụnyere omenala cell, anụ ahụ, mmiri ahụ, wdg na ihe karịrị 7,000 mRNA-Seq ọrụ mechiri na-ekpuchi mpaghara nyocha dị iche iche.

ØSistemụ njikwa mma siri ike: Isi njikwa mma isi site na usoro niile gụnyere nkwadebe nlele, nkwadebe ọbá akwụkwọ, usoro na bioinformatics ka a na-enyocha ya nke ọma iji wepụta nsonaazụ dị elu.

ØOtutu ọdụ data dị maka nkọwa ọrụ yana ọmụmụ ịba ụba iji mezuo ebumnuche nyocha dị iche iche.

ØỌrụ ire ere: Ọrụ ire ere gachara maka ọnwa 3 ka arụchara ọrụ, gụnyere nlebanya ọrụ, mgbapụ nsogbu, nsonaazụ Q&A, wdg.

Ihe nlele chọrọ na nnyefe

Ụlọ akwụkwọ	Usoro usoro	akwadoro data	Njikwa ogo
Poly A bara ụba	Ilumina PE150	6 gb	Q30≥85%

Ihe atụ chọrọ:

Nucleotides:

Ịdị ọcha	Iguzosi ike n'ezi ihe	Ọnụ ego
OD260/280≥1.7-2.5 OD260/230≥0.5-2.5Enwere oke ma ọ bụ enweghị protein ma ọ bụ mmetọ DNA egosiri na gel.	Maka osisi: RIN≥6.5; Maka anụmanụ: RIN≥7; 28S/18S≥1.0; nwere oke ma ọ bụ enweghị mbuli elu.	Conc.≥30 ng/μl;Mpịakọta ≥ 10 μl;Mkpokọta ≥ 1.5 μg

Anụ ahụ: Ibu (akọrọ):≥1 g
* Maka anụ ahụ dị ntakịrị karịa 5 mg, anyị na-akwado ka izipu ihe nlele anụ ahụ kpọnwụrụ akpọnwụ (na mmiri mmiri nitrogen).

Nkwụsị cell:Ọnụ ọgụgụ cell = 3×10⁶- 1 × 10⁷
* Anyị na-akwado ibupu cell lysate oyi kpọnwụrụ.Ọ bụrụ na ọnụ ọgụgụ cell ahụ pere mpe karịa 5 × 105, a na-atụ aro ka oyi kpọnwụrụ na nitrogen mmiri mmiri, nke ka mma maka mmịpụta micro.

Ụdị ọbara:Olu ≥1 ml

Microorganism:Uka ≥ 1 g

Nnyefe nlele akwadoro

Akpa: 2 ml centrifuge tube (Tin foil adịghị akwadoro)

Nkado nlere anya: Otu+ megharịa dịka A1, A2, A3;B1, B2, B3......

Mbupu:

Ikpụrụ kpọrọ nkụ: Ọ dị mkpa ka a kwakọba ihe nlele n'ime akpa wee lie ya na ice kpọrọ nkụ.
Ọkụ RNAstable: Enwere ike ịkpọsa sample RNA na tube stabilization RNA (dịka ọmụmaatụ RNAstable®) ma bufee ya n'ime ụlọ okpomọkụ.