Iriburiro:

RNA, izina ry'ubumenyi bwa ribonucleic aside, mu magambo ahinnye akomoka kuri Acide ya RiboNucleic, ni ubwoko bwa acide nucleique bwakozwe mu guhuza byibuze na mirongo ya ribonucleotide binyuze mu mitsi ya fosifori.

Intego yo gushakisha RNA nugukuramo RNA Yuzuye idafite umwanda, kandi ihame ryibanze ryo gukuramo RNA rishobora kuvugwa muri make kuburyo bukurikira:

1. Banza uhagarike selile

2. Noneho ukureho umwanda wa macromolecular (proteyine, lipide, ADN, nibindi)

3. Hanyuma, RNA iragwa mugihe ikuyemo umunyu hamwe numuti ukomoka kumubiri, no kurushaho kweza RNA

Uburyo bwashize nubu bwo gukuramo

Igisekuru cya mbere cyuburyo bwo gukuramo RNA nuburyo bwa TRIzol bwimvura (reagent ikoreshwa harimo guanidine isothiocyanate, phenol, chloroform, nibindi) yahimbwe na sogokuru witwa Piotr Chomczynski na nyirakuru Nicoletta Sacchi, kuko iki gihangano cyemerera gukuramo RNA byihuse kandi byoroshye, kuva icyo gihe byatangiye kuzamuka mubushakashatsi bwa RNA.

Nyuma, Sogokuru yagurishije iyi patenti kuri Invitrogen, maze Invitrogen iteza imbere ubu buryo ku isi kandi ihinduka uburyo bukoreshwa cyane.

Nyuma, Invitrogen yaguzwe na Thermo!

Ibicuruzwa byerekana: Invitrogen ya TRIzol reagent

Nyamara, uburyo bwimvura ya TRIzol busaba imvura nyinshi nogukuramo RNA, kandi reagent zikoreshwa nuburozi kandi bifata igihe kinini kugirango zikore.

Kubwibyo, igisekuru cya kabiri inkingi-enzyme yo kweza yabayeho, ikoresheje silika gel membrane nkibikoresho byihariye bya acide nucleic aside adsorption kugirango bigarure cyane RNA mu cyitegererezo mugihe ikuraho ibindi byanduye.

Uburyo bwa kabiri bwibisekuru byoroshya imikorere hashingiwe kumikorere ihamye, bigabanya ingaruka ziterwa, kandi bitezimbere imikorere yo gukuramo RNA.

Ibicuruzwa byerekana: QIAGEN RNeasy nibicuruzwa byinshi murugo

Nyamara, uburyo bwombi bwambere nubwa kabiri busaba gukoresha DNase kugirango ukureho kwanduza ADN, bizana bimwe mubitagenda neza mubushakashatsi bwakurikiyeho.DNase isigaye izagabanya imikorere yo kwandukura no kugabanya umurongo wa mbere wa cDNA synthesis, bityo bigire ingaruka kubisubizo byukuri.

Kubwibyo, uburyo bwa gatatu bwo kuvoma uburyo butezimbere hashingiwe ku gisekuru cya kabiri.Ubushakashatsi ntibukeneye kongeramo imisemburo iyo ari yo yose, kandi RNA irashobora kuboneka byoroshye hamwe ninkingi ebyiri gusa.

Inkingi ya 1: ADN-Isukura Inkingi kugirango isibe ADN

Inkingi ya 2: RNA-Inkingi gusa Adorbs kandi igarura RNA

Igikorwa cyoroshye ntabwo gikiza umwanya gusa, ariko kandi cyoroshya imikorere yubushakashatsi bwo hasi!

Ibicuruzwa byerekana: QIAGEN RNeasy Yongeyeho naForegeneIbicuruzwa bya RNA byoza

（RNeasy Plus）

（Foregene'sGutera Igiteranyo Cyuzuye cya RNA）

Nkubwire utuje!Ibikoresho bya RNA bya Foregene birerekana kandi byoroshye gukoresha RNase Eraser, ishobora gukuraho burundu 400μg ya RNase hejuru yumunota 1, nta kongeramo ibintu byuburozi mumubiri wumuntu!

Pic1: Imbonerahamwe yo kugereranya ibikorwa

Ibicuruzwa bya Foregene birashobora kurangiza gukuramo RNA mu ndirimbo ebyiri gusa, ntugatume abandi bagirira ishyari iterambere ryawe!

Pic2:Eibisubizo byubushakashatsi imbonerahamwe yo kugereranya

Igisekuru cya mbere (1: TRIzol)

Igisekuru cya kabiri (2: RNeasy ya QIAGEN)

Igisekuru cya gatatu (3: RNIasy ya QIAGEN wongeyeho, 4: Foregene)

Pic3: igishushanyo cya qPCR (igishushanyo mbonera cya amplification)

Igisekuru cya mbere (icyatsi bisobanura TRIzol -, umutuku bisobanura TRIzol +)

Igisekuru cya kabiri (ikirere cyubururu bisobanura QIAGEN RNeasy-, umutuku bisobanura QIAGEN RNeasy +)

Igisekuru cya gatatu (ubururu bisobanura QIAGEN RNeasy wongeyeho, icyatsi bisobanura Foregene)

.

Buri kuzamura tekinike bivuze ko imikorere yikigeragezo itera imbere muburyo bworoshye kandi bworoshye kubakoresha.

Foregene igamije gufasha abashakashatsi benshi ba siyansi kuzigama igihe nigiciro, kubona ibisubizo byubushakashatsi bufite ireme, no gufatanya gukora ubushakashatsi bwubumenyi bufite agaciro.