China Eukaryotic mRNA sequencing-Illumina fektheri le bahlahisi |BMK

Eukaryotic mRNA sequencing-Illumina

tatellano ea mRNA e thusa ho hlahisa li-mRNA tsohle tse ngotsoeng ho tsoa liseleng tlas'a maemo a itseng.Ke thekenoloji e matla ea ho senola boemo ba liphatsa tsa lefutso, mekhoa ea liphatsa tsa lefutso le mekhoa ea limolek'hule ea mekhoa e itseng ea likokoana-hloko.Ho fihla hajoale, tatellano ea mRNA e sebelisitsoe haholo lipatlisisong tsa mantlha, tlhahlobo ea bongaka, nts'etsopele ea lithethefatsi, jj.

Sethala: Illumina NovaSeq 6000

Iteanye le rona

Lintlha tsa Tšebeletso

Bioinformatics

Demo Results

Melemo

ØBa nang le Boiphihlelo bo Haholo: Ho se ho sebelitsoe lisampole tse fetang 200,000 ho BMK e koahelang mefuta e fapaneng ea lisampole, ho kenyeletsoa setso sa lisele, lisele, mokelikeli oa 'mele, joalo-joalo le merero e fetang 7,000 ea mRNA-Seq e koetsoeng e koahelang libaka tse fapaneng tsa lipatlisiso.

ØSistimi e thata ea taolo ea boleng: Lintlha tsa taolo ea boleng ba mantlha ka mehato eohle ho kenyeletsoa tokiso ea lisampole, tokiso ea laeborari, tatelano le bioinformatics li tlas'a leihlo le nchocho e le ho fana ka liphetho tsa boleng bo holimo.

ØMananeo a mangata a fumanehang bakeng sa litlhaloso tsa ts'ebetso le lithuto tsa ntlafatso ho phethahatsa lipheo tse fapaneng tsa lipatlisiso.

ØLitšebeletso tsa ka mor'a thekiso: Litšebeletso tsa ka mor'a thekiso li sebetsa likhoeli tse 3 ha projeke e phetheloa, ho kenyelletsa le tatellano ea merero, ho rarolla mathata, liphetho tsa Q&A, jj.

Litlhoko tsa Mohlala le Phano

Laebrari	Leano la tatellano	Lintlha li khothalelitsoe	Taolo ea Boleng
Poly A matlafatsa	Illumina PE150	6 Gb	Q30≥85%

Litlhoko tsa Mohlala:

Li-nucleotides:

Bohloeki	Botšepehi	Chelete
OD260/280≥1.7-2.5 OD260/230≥0.5-2.5Tšoaetso e fokolang kapa ha e na protheine kapa DNA e bontšitsoeng ho gel.	Bakeng sa limela: RIN≥6.5; Bakeng sa liphoofolo: RIN≥7;28S/18S≥1.0; bophahamo bo lekanyelitsoeng kapa bo se nang motheo	Conc.≥30 ng/μl;Molumo ≥ 10 μl;Kakaretso ≥ 1,5 μg

Tila: Boima ba 'mele (o omileng):≥1 g
*Bakeng sa linama tse nyane ho feta 5 mg, re khothaletsa ho romella sampuli ea lithane tsa "flash" tse leqhoa (ka naetrojene e metsi).

Ho emisoa ha lisele:Palo ea lisele = 3×10⁶-1 × 10⁷
* Re khothaletsa ho romella li-lysate tse leqhoa.Haeba sele eo e le nyane ho feta 5 × 105, ho khothaletsoa flash e hoamisitsoeng ka har'a metsi a naetrojene, e leng khetho e ntle bakeng sa ho ntšoa ha nyane.

Mehlala ea mali:Bophahamo ≥1 ml

Microorganism:Boima ≥ 1 g

Tlhahiso ea Mehlala e khothalelitsoeng

Setshelo: 2 ml centrifuge tube (Tin foil ha e khothalletsoe)

Mohlala oa ho ngola: Sehlopha+katiso mohlala A1, A2, A3;B1, B2, B3....

Thomello:

Leqhoa le omileng: lisampole li hloka ho kenngoa ka mekotleng ebe li patoa ka leqhoa le omileng.
Li-tubes tsa RNAstable: lisampole tsa RNA li ka omisoa ka har'a li-tube tsa RNA stabilization (mohlala, RNAstable®) ebe li romelloa ka mocheso oa kamore.