Lihlahisoa

Eukaryotic mRNA Sequencing-Illumina

tatellano ea mRNA e nolofalletsa profil ea 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 joale, tatellano ea mRNA e sebelisitsoe haholo lipatlisisong tsa mantlha, tlhahlobo ea bongaka, nts'etsopele ea lithethefatsi, jj.

Sethala: Illumina NovaSeq Platform

Iteanye le rona

Lintlha tsa Tšebeletso

Bioinformatics

Demo Results

Melemo

● Ba nang le Boiphihlelo bo Haholo: Ho entsoe lisampole tse fetang 200,000 ka har'a 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 mRNA-Seq e koetsoeng e koahelang libaka tse fapaneng tsa lipatlisiso.

● Sistimi e thata ea taolo ea boleng: Lintlha tsa taolo ea boleng ba mantlha ka mehato eohle e kenyelletsang ho lokisoa ha sampuli, ho lokisoa laebraring, tatelano le bioinformatics li tlas'a leihlo le nchocho e le ho fana ka liphetho tsa boleng bo holimo.

● Mananeo a mangata a teng bakeng sa litlhaloso tsa tšebetso le lithuto tsa ntlafatso ho phethahatsa lipakane tse fapaneng tsa lipatlisiso.

● Litšebeletso tsa ka mor'a thekiso: Litšebeletso tsa ka mor'a thekiso li sebetsa likhoeli tse 3 ha morero o phethoa, ho kenyelletsa le ho latela merero, ho rarolla mathata, liphetho tsa Q&A, joalo-joalo.

Litlhoko tsa Mohlala le Phano

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

Litlhoko tsa Mohlala:

Li-nucleotides:

Conc.(ng/μl)

Chelete (μg)

Bohloeki

Botšepehi

≥20

≥ 0.5

OD260/280=1.7-2.5

OD260/230=0.5-2.5

Tšoaetso e fokolang kapa ha e na protheine kapa DNA e bontšitsoeng ho gel.

Bakeng sa limela: RIN≥6.5;

Bakeng sa liphoofolo: RIN≥7.0;

5.0≥28S/18S≥1.0;

bophahamo bo fokolang kapa bo se nang motheo

Tila: Boima ba 'mele (o omileng):≥1 g
*Bakeng sa linama tse nyane ho feta 5 mg, re khothaletsa ho romela sampole ea lithane tse hatselitsoeng (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.

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.