Kurandura inama za pipette hamwe na EP tubes, nibindi.

1. Tegura 0.1% (igihumbi) DEPC (ibintu bifite ubumara bukabije) n'amazi ya deioniyo, uyikoreshe witonze mumashanyarazi, hanyuma ubibike kuri 4 ° C kure yumucyo;

Amazi ya DEPC ni amazi meza atunganywa na DEPC kandi akayungurura ubushyuhe bwinshi n'umuvuduko mwinshi.Bipimishije kubusa RNase, DNase na proteinase.

2. Shira umuyoboro wa pipette na EP muri 0.1% DEPC, hanyuma urebe ko umuyoboro wa pipette na EP wuzuye 0.1% DEP.

3. Kurinda urumuri, reka guhagarara, ijoro ryose (12-24h)

4. Agasanduku karimo inama na EP tube ntigomba gushirwa muri DEPC.Nyuma yo gukuramo hafi amazi ya DEPC mumutwe cyangwa EP tube, bipakira hanyuma ubizenguruke.

5. dogere selisiyusi 121, 30min

6. dogere selisiyusi 180, yumisha amasaha menshi (byibuze amasaha 3)

Icyitonderwa: a.Wambare uturindantoki twa latex na masike mugihe ukora DEPC!b, cyangwa idafite DEPC sterilisation, 130 ℃, 90min autoclave (laboratoire nyinshi ubushyuhe bwo hejuru bwa kabiri)

Ibitekerezo byo gukuramo RNA

Ibintu bibiri byingenzi bya tissue RNA kwigunga

Kwangirika kwa RNA n'ibisigisigi byanduye mumyenda,kubyerekeranye no gutesha agaciro, reka tubanze turebe impamvu RNA yakuwe mu ngirabuzimafatizo zidafite umuco.Gukuramo RNA iriho reagent zose zirimo ibice bibuza RNase byihuse.Ongeramo lysate mu ngirabuzimafatizo zifite umuco, hanyuma ubivange gusa, selile zose zirashobora kuvangwa neza na lysate, kandi selile zuzuye.Ingirabuzimafatizo zimaze guterwa, ibikoresho bikora muri lysate bihita bibuza RNase idasanzwe, bityo RNA ikomeza kuba ntamakemwa.Nukuvuga ko, kubera ko ingirabuzimafatizo zifite imico zoroshye kandi zihuye neza na lysate, RNA yabo ntabwo yangirika byoroshye;kurundi ruhande, RNA mumyanya yangiritse byoroshye kuko selile zo mumyanya ntizoroshye guhura na lysate.kubera guhuza bihagije.Noneho,dufate ko hari uburyo bwo guhindura tissue ingirabuzimafatizo imwe mugihe uhagarika ibikorwa bya RNA, ikibazo cyo gutesha agaciro gishobora gukemuka rwose.

Gusya kwa azote ni uburyo bwiza cyane nkubwo.Nyamara, uburyo bwo gusya bwa azote bwamazi buteye ikibazo cyane cyane iyo umubare wintangarugero ari munini.Ibi byabyaye ikintu cyiza gikurikira: homogenizer.Uwitekahomogenizeruburyo ntibusuzuma ikibazo cyukuntu ibikorwa bya RNase bibujijwe mbere yuko selile zihura na lysate, ahubwo igasenga ngo igipimo cyo guhagarika ingirangingo cyihuta kuruta igipimo RNase yo mu nda itesha agaciro RN.

Ingaruka ya homogenizer yamashanyarazi nibyiza,n'ingaruka z'ikirahuri homogenizer ni mbi, ariko muri rusange, uburyo bwa homogenizer ntibushobora gukumira ibintu bitesha agaciro.Kubwibyo, niba gukuramo byangiritse, amashanyarazi yumwimerere agomba gukoreshwa mugusya hamwe na azote yuzuye;ikirahure cyumwimerere homogenizer igomba guhinduka kuri homogenizer yamashanyarazi cyangwa igasya neza na azote yuzuye.Ikibazo kirashoboka 100%.bikemuke.

Ikibazo gisigaye cyanduye kigira ingaruka kubushakashatsi bwakurikiyeho gifite impamvu zitandukanye kuruta gutesha agaciro, kandi ibisubizo biratandukanye.Mu gusoza,niba hari gutesha agaciro cyangwa umwanda usigaye muri tissue, uburyo bwo kuvoma / reagent kubintu byihariye byubushakashatsi bigomba kuba byiza.Ntugomba gukoresha ingero zawe z'agaciro kugirango utezimbere: urashobora kugura inyamaswa ntoya nk'amafi / inkoko ku isoko, ugafata igice kijyanye n'ibikoresho byo gukuramo RNA, ikindi gice cyo gukuramo poroteyine - gusya ukoresheje umunwa, igifu n'amara Gukuramo.

Intego RNA ya RNA yakuweho ikoreshwa mubushakashatsi butandukanye bwo gukurikirana, kandi ubuziranenge bwabwo buratandukanye

kubaka isomero rya cDNA bisaba ubunyangamugayo bwa RNA nta bisigisigi bya enzyme reaction;Amajyaruguru arasaba ubunyangamugayo bwa RNA nibisabwa hasi kugirango enzyme reaction ibuza ibisigisigi;RT-PCR ntabwo isaba ubunyangamugayo bwa RNA cyane,ariko irabuza reaction ya enzyme.Ibisabwa bisigaye birakomeye.Iyinjiza igena ibisohoka;igihe cyose intego ni ukubona isuku ihanitse RNA, bizatwara abantu namafaranga.

Gukusanya / Kubika Ingero

Ibintu bigira ingaruka mbi ku kwangirika Nyuma yicyitegererezo kiva mumubiri muzima / cyangwa ibidukikije byambere bikura, imisemburo ya endogenous muri sample izatangira gutesha agaciro RNA,kandi igipimo cyo gutesha agaciro kijyanye nibirimo imisemburo ya endogenous n'ubushyuhe.Ubusanzwe, hari inzira ebyiri gusa zo guhagarika burundu ibikorwa bya enzyme ya endogenous: ongeramo lysate ako kanya hanyuma homogenize neza kandi byihuse;gabanya uduce duto hanyuma uhite uhagarika muri azote yuzuye.Ubwo buryo bwombi busaba gukora byihuse.Iyanyuma irakwiriye kuburugero rwose, mugihe iyambere ikwiranye gusa nuduce dufite ibintu bike bigize selile na enzymes za endogenous kandi byoroshye guhuza abantu.By'umwihariko, ibimera, umwijima, thymus, pancreas, impyiko, ubwonko, ibinure, imitsi, nibindi bikonjeshwa neza na azote yuzuye mbere yo gukomeza.

Gucamo ibice no guhuza ingero

Ibintu bigira ingaruka ku gutesha agaciro no gutanga umusaruro Icyitegererezo nikuri homogenisation yuzuye, aribyo kurekura byuzuye kandi byuzuye bya RNA.Ingirabuzimafatizo zirashobora guhurizwa hamwe bitavunitse.Tissue irashobora guhuzwa gusa nyuma yo kumeneka.Umusemburo na bagiteri bigomba gucika hamwe na enzymes zijyanye nabyo mbere yuko bihuzwa.Uturemangingo dufite enzyme yo hepfo ya endogenous kandi byoroshye homogenisation irashobora guhonyorwa no guhuriza hamwe icyarimwe muri lysate na homogenizer;ibimera, umwijima, thymus, pancreas, intanga, ubwonko, ibinure, ingirangingo nizindi ngero, Zirashobora kuba nyinshi mumisemburo ya endogenous cyangwa ntabwo byoroshye kubana,guhagarika tissue rero na homogenisation bigomba gukorwa bitandukanye.Uburyo bwizewe kandi butanga umusaruro cyane bwo gucamo ibice ni ugusya hamwe na azote yuzuye, kandi uburyo bwizewe bwo guhuza ibitsina ni ugukoresha amashanyarazi.Icyitonderwa kidasanzwe kijyanye no gusya hamwe na azote yuzuye: icyitegererezo ntigomba gukonjeshwa mugihe cyose cyo gusya, kuko imisemburo ya endogenous ishobora gukora cyane iyo ikonje.

Guhitamo lysate

Ingaruka zorohereza imikorere hamwe nimpamvu ziterwa na endogenous zanduye zisigaye zikoreshwa cyane lysis ibisubizo birashobora guhagarika ibikorwa bya RNase.Kubwibyo, ingingo yingenzi yo guhitamo igisubizo cya lysis ni ukuzirikana hamwe nuburyo bwo kweza.Hariho ikintu kimwe kidasanzwe:ingero zifite enzyme nyinshi ya endogenous irasabwa gukoresha lysate irimo fenol kugirango yongere ubushobozi bwo kudakora imisemburo ya endogenous.

Guhitamo uburyo bwo kweza

Ibintu bigira ingaruka kumyanda isigaye ya endogenous, umuvuduko wo gukuramo Kuburugero rwiza nkutugingo ngengabuzima, ibisubizo bishimishije birashobora kuboneka hamwe nuburyo ubwo aribwo bwose bwo kweza buri hafi.Ariko kubindi byitegererezo byinshi, cyane cyane abafite umwanda mwinshi nkibimera, umwijima, bagiteri, nibindi, guhitamo uburyo bukwiye bwo kweza ni ngombwa.Inkingi ya centrifugal yo kweza ifite umuvuduko wo gukuramo byihuse kandi irashobora gukuraho neza umwanda ugira ingaruka kumyitwarire ya nyuma ya RNA, ariko irazimvye (Foregene irashobora gutanga ibikoresho bikoresha amafaranga menshi, ibisobanuro birambuye kandahano);ukoresheje uburyo bwubukungu nubusanzwe bwo kweza, nkimvura ya LiCl, irashobora kandi kubona ibisubizo bishimishije, ariko igihe cyo gukora ni kirekire..

"Disipuline eshatu nuburyo umunani" zo gukuramo RNA

Indero 1:Shira iherezo ku kwanduza imisemburo ya exogenous.

Icyitonderwa 1:Wambare cyane masike na gants.

Icyitonderwa 2:Imiyoboro ya centrifuge, imitwe yinama, inkoni ya pipeti, ibigega bya electrophoreis, nintebe zigeragezwa zigira uruhare mubigeragezo bigomba kujugunywa neza.

Icyitonderwa 3:Reagents / ibisubizo bigira uruhare mubigeragezo, cyane cyane amazi, bigomba kuba bidafite RNase.

Indero 2:Hagarika ibikorwa bya enzymes endogenous

Icyitonderwa 4:Hitamo uburyo bukwiye bwo guhuza ibitsina.

Icyitonderwa 5:Hitamo lysate ikwiye.

Icyitonderwa 6:Kugenzura umubare wintangiriro yicyitegererezo.

Indero ya 3:Sobanura intego yawe yo gukuramo

Icyitonderwa 7:Hamwe na sisitemu ya lysate yegera umubare ntarengwa wo gutangira icyitegererezo, igipimo cyo gutsinda cyagabanutse cyane.

Icyitonderwa 8:Igipimo cyubukungu cyonyine cyo gukuramo RNA neza ni intsinzi mubushakashatsi bwakurikiyeho, ntabwo itanga umusaruro.

Isoko 10 yambere yo kwanduza RNase

1. Urutoki nisoko yambere yimisemburo ya exogenous, bityo gants igomba kwambara kandi igasimburwa kenshi.Byongeye kandi, masike nayo igomba kwambarwa, kuko guhumeka nabyo ni isoko yingenzi ya enzymes.Inyungu yinyongera yo kwambara mask ya gants ni ukurinda uwagerageje.

2. Impanuro za pipette, imiyoboro ya centrifuge, imiyoboro - RNase ntishobora gukoreshwa na sterisizione yonyine, bityo inama za pipette hamwe nigituba cya centrifuge bigomba kuvurwa na DEPC, kabone niyo byashyizweho ikimenyetso nka DEPC ivurwa.Nibyiza gukoresha umuyoboro udasanzwe-wihariye, uhanagure umupira wa pamba ya alcool 75% mbere yo gukoresha, cyane cyane inkoni;wongeyeho, menya neza ko udakoresha gukuramo umutwe.

3. Amazi / buffer agomba kuba adafite umwanda wa RNase.

4. Nibura ameza yikizamini agomba guhanagurwa neza hamwe nudupapuro twa pamba ya alcool 75%.

5.Endogenous RNase Uturemangingo twose turimo imisemburo ya endogenous, bityo gukonjesha vuba ingirangingo hamwe na azote yuzuye ni inzira nziza yo kugabanya kwangirika.Amazi ya azote yo kubika / gusya mubyukuri ntabwo byoroshye, ariko niyo nzira yonyine yimyenda ifite urwego rwinshi rwa enzymes ya endogenous.

6. Icyitegererezo cya RNA Ibicuruzwa bivamo RNA bishobora kuba birimo ibimenyetso byanduye RNase.

7. Gukuramo plasmide Gukuramo plasmid akenshi bikoresha Rnase kugirango bitesha agaciro RNA, kandi Rnase isigaye igomba guhishwa na Proteinase K hanyuma igakurwa na PCI.

8. Ububiko bwa RNA Nubwo bwabitswe ku bushyuhe buke, urugero rwa RNase ruzatera kwangirika kwa RNA.Igisubizo cyiza cyo kubungabunga RNA igihe kirekire ni uguhagarika umunyu / inzoga, kuko inzoga zibuza ibikorwa byose byimisemburo mubushyuhe buke.

9. Iyo cations (Ca, Mg) zirimo izo ion, gushyushya kuri 80C muminota 5 bizatera RNA kumeneka, niba rero RNA ikeneye gushyuha, igisubizo cyo kubungabunga gikeneye kubamo chelating (1mM Sodium Citrate, pH 6.4).

10. Enzymes zikoreshwa mubushakashatsi bwakurikiyeho zirashobora kwanduzwa na RNase.

Inama 10 zo gukuramo RNA

1: Irinde vuba ibikorwa bya RNase.Ingero zahagaritswe vuba nyuma yo gukusanya, kandi RNase idakorwa nigikorwa cyihuse mugihe cya lysis.

2: Hitamo uburyo bukwiye bwo gukuramo tissue zifite ibinini byinshi bya ribozyme, kandi tipusi ya adipose nibyiza gukoresha uburyo burimo fenol.

3: Ubwiza bwo guhanura busaba Amajyaruguru, cDNA kubaka isomero bisaba ubunyangamugayo buhanitse, kandi RT-PCR na RPA (Ribonuclease protection assay) ntibisaba ubunyangamugayo buhanitse.RT-PCR isaba ubuziranenge bwinshi (ibisigisigi bya enzyme inhibitor).

4: Guhuza byimazeyo ni urufunguzo rwo kuzamura umusaruro no kugabanya iyangirika.

5: Reba ubunyangamugayo bwa RNA electrophoresis detection, 28S: 18S = 2: 1 nikimenyetso cyuzuye, 1: 1 nayo iremewe kubushakashatsi bwinshi.

6: Gukuraho ADN kuri RT-PCR, isesengura ryibisobanuro Nibyiza gukoresha Dnase I kugirango ukure ADN.

7: Kugabanya kwanduza imisemburo ya exogenous - enzymes ntishobora gutumizwa hanze.

8: Iyo ushizemo aside nucleic-acide nkeya, hagomba kongerwamo reagent ya co-imvura.Ariko kugirango wirinde co-precitant irimo enzymes hamwe na ADN yanduye.

9: Kuraho neza RNA, nibiba ngombwa, shyushya kuri 65C muminota 5.

uburyo bukwiye bwo kubika

Irashobora kubikwa kuri –20C mugihe gito, no kuri 80C igihe kirekire.Intambwe yambere mugutezimbere umusaruro wa RNA nukumenya ko RNA yibigize ingero zitandukanye zitandukanye cyane.Ubwinshi bwinshi (2-4ug / mg) nk'umwijima, pancreas, umutima, ubwinshi bwo hagati (0.05-2ug / mg) nk'ubwonko, urusoro, impyiko, ibihaha, thymus, ovary, ubwinshi (<0.05ug / mg) mg) nk'uruhago, amagufa, ibinure.

1: Lyse selile kurekura RN - niba RNA itarekuwe, umusaruro uzagabanuka.Amashanyarazi ya homogenisation akora neza kurenza ubundi buryo bwo guhuza ibitsina, ariko birashobora no gukenera guhuzwa nubundi buryo, nka mashing ya azote yuzuye, igogorwa ryimisemburo (Lysozyme / Lyticase)

2: Gukwirakwiza uburyo bwo kuvoma.Ibibazo bikomeye hamwe nuburyo bushingiye kuri fenolike ni stratifike ituzuye hamwe no gutakaza igice cya RNA (ndengakamere ntishobora kuvaho burundu).Gutondekanya kutuzuye biterwa na aside nucleic nyinshi hamwe na proteyine nyinshi, zishobora gukemurwa no kongera urugero rwa lysate yakoreshejwe cyangwa kugabanya urugero rwicyitegererezo.Intambwe yo gukuramo chloroform yongewe kumubiri wa adipose.Igihombo cya RNA kirashobora kugabanuka mugupompa inyuma cyangwa mugukuraho urwego kama rukurikirwa na centrifugation.Ikibazo kinini hamwe ninkingi centrifugation ishingiye kuburyo burenze urugero.

Inama zo gukuramo ibintu bya kera

1. Isuku ya Fenol: Ongeramo ingano ingana na 1: 1 Fenol / Chloroform hanyuma uvange cyane muminota 1-2.Centrifuge kumuvuduko mwinshi muminota 2.Witonze ukureho ndengakamere (80-90%).Ntuzigere ugera kumurongo wo hagati.Ingano ingana nigisubizo cya reaction irashobora kongerwa kuri Phenol / Chloroform hanyuma ikurwaho ndengakamere.Ibintu ndengakamere byombi birashobora kuvangwa hamwe kugirango imvura igwa acide nucleic kugirango umusaruro wiyongere.Ntukitonda cyane mugihe uvanze, kandi ntugerageze gukuraho ibintu ndengakamere.

2. Gukaraba hamwe na 70-80% Ethanol: Mugihe cyo gukaraba, aside nucleic igomba guhagarikwa kugirango umunyu usigaye woge.Mugihe kimwe, ako kanya nyuma yo gusuka Ethanol, centrifuge kumuvuduko mwinshi mumasegonda make, hanyuma ukureho Ethanol isigaye hamwe na pipeti.Gabanya nyuma yo guhagarara mubushyuhe bwicyumba muminota 5-10.

11. Gukuramo amashyirahamwe adasanzwe

1. Fibrous tissue: Urufunguzo rwo gukuramo RNA mumitsi ya fibrous nkumutima / imitsi ya skeletale ni uguhagarika rwose ingirangingo.Izi nyama zifite ubwinshi bwimikorere ya selile, kubwibyo ingano ya RNA kuburemere bwibice bya tissue iba mike, kandi nibyiza gukoresha amafaranga menshi yo gutangira bishoboka.Witondere gusya neza neza mugihe gikonje.

2. Ibice birimo proteine ​​nyinshi / ibinure: ubwonko / ibinure byimboga ni byinshi.Nyuma yo gukuramo PCI, ndengakamere irimo floccules yera.Indengakamere igomba kongera gukururwa na chloroform.

3. Uturemangingo turimo aside nucleic nyinshi / ribozyme: spleen / thymus ifite aside nucleic aside nyinshi hamwe na ribozyme.Gusya ingirangingo mubihe bikonje bikurikirwa na homogenisation yihuse birashobora gukora neza ribozymes.Ariko, niba lysate igaragara cyane (kubera aside irike ya nucleique), gukuramo PCI ntibishobora gutondeka neza;kongeraho lysate irashobora gukemura iki kibazo.Gukuramo PCI nyinshi birashobora gukuraho ADN zisigaye.Niba imvura yera igize ako kanya nyuma yo kongeramo inzoga, byerekana kwandura ADN.Kongera gukuramo PCI acide nyuma yo guseswa birashobora gukuraho ADN yanduye.

4. Ibimera by'ibihingwa: Ibimera by'ibihingwa biragoye kuruta inyamaswa.Mubisanzwe, ibimera biri mubutaka bwa azote, bityo kwangirika kwa RNA na enzymes za endogenous ntibisanzwe.Niba ikibazo cyo gutesha agaciro kidakemutse, byanze bikunze biterwa numwanda uri murugero.Umwanda ukubiye mubihingwa byinshi bizaganisha ku bisigazwa, kandi impamvu yibisigara ni ukubera ko iyo myanda ifite aho ihuriye na RNA: uragusha kandi nanjye ndagusha, kandi nawe adsorb na I adsorb.Ibiranga byerekana ko ari inzitizi zikomeye za enzyme.

Kugeza ubu, ibicuruzwa biva mu bucuruzi bya RNA birashobora guhuzwa n’inyama zose z’inyamanswa hamwe n’ibihinduka bito, ariko hariho reagent nke zo gukuramo RNA zishobora kuba zikwiranye ningingo nyinshi z’ibimera.Kubwamahirwe, Foregene irashobora gutanga umwiharikoibihingwa byo gukuramo RNA, dufiteGutera Igiteranyo Cyuzuye RNA, Tera Igiteranyo cya RNA Igikoresho cyongeyeho.Iyanyuma yagenewe byumwihariko kubimera bifite polysaccharide nyinshi hamwe na polifenol.Kubikuramo RNA, ibitekerezo byabakoresha laboratoire nibyiza cyane.

12. Ingaruka zo gukonjesha no gukonjesha Icyitegererezo cyakonje gishobora kuba kinini, kandi kigomba gucibwa mbere yo gukoreshwa mu gukuramo RNA.Ingero zikunda gushonga (birashoboka igice) mugihe cyo gutema.Ingero zikonje zishobora gukenera gupimwa mbere yo gukuramo RNA, kandi gukonjesha bizabaho muriki gihe.Rimwe na rimwe, gukonjesha icyitegererezo nabyo bibaho mugihe cyo gusya azote;cyangwa icyitegererezo cyakonjeshejwe cyongewe kuri lysate nta gusya kwa azote yuzuye, kandi gukonjesha bizabaho mbere yuko homogenisation yuzuye.Ubushakashatsi bwerekanye ko ingirabuzimafatizo zafunzwe zikunze kwangirika kwa RNA mugihe cyo gukonja kuruta imyenda mishya.Impamvu ishobora kuba: Gukonjesha-guhagarika ibikorwa bihagarika imiterere muri selire, bigatuma byoroha imisemburo ya endogenous guhura neza na RNA.

13. Gucira ubuziranenge bwa RNA Mubisanzwe, electrophoreis ikoreshwa mugucira urubanza ubusugire bwa RNA, naho A260 / A280 ikoreshwa mugucira isuku RNA.Mubyigisho, RNA idahwitse ifite igipimo cya 28S: 18S = 2.7: 1, kandi amakuru menshi ashimangira igipimo cya 28S: 18S = 2: 1.Ikigaragara ni uko hafi ya RNA yakuwe mu ngero zitari selile ziri mu kigereranyo cya 2: 1 (ibi byabonetse hakoreshejwe Agilent Bioanalyser).

Ibisubizo bya electrophoreis ya RNA bigira ingaruka kubintu byinshi, harimo imiterere ya kabiri, imiterere ya electrophoreis, umutwaro wikitegererezo, urugero rwuzuye rwa EB, nibindi. Koresha amashanyarazi kavukire kugirango umenye RNA kandi ukoreshe ADN Marker nkigenzura.Niba 28S kuri 2kb na 18S kuri 0.9kb isobanutse, na 28S: 18S> 1, ubunyangamugayo burashobora kuzuza ibisabwa mubushakashatsi bwakurikiyeho.

A260 / A280 ni ikimenyetso cyateje urujijo rwinshi.Mbere ya byose, birakenewe gusobanura ibisobanuro byumwimerere byiki kimenyetso cya acide nucleic: RNA yera, A260 / 280 = hafi 2.0.RNA yera n '' impamvu 'na A260 / A280 = 2 ni' ingaruka '.Ubu abantu bose bakoresha A260 / A280 nk '' impamvu ', batekereza ko "niba A260 / A280 = 2, noneho RNA ni nziza", mubisanzwe bitera urujijo.

Niba ubishaka, urashobora kongeramo reagent nkeya ikoreshwa mugukuramo, nka fenol, guanidine isothiocyanate, PEG, nibindi, kurugero rwa RNA, hanyuma ugapima igipimo cya A260 / A280.Ikigaragara ni uko ibyinshi mu reagent zikoreshwa mu gukuramo RNA, kimwe n’umwanda mwinshi uri muri sample, bikurura A260 na A280, bikagira ingaruka kuri A260 / A280.

Uburyo bwigisha cyane kurubu ni ugusuzuma ibyitegererezo bya RNA murwego rwa 200-300 nm.Umurongo wa RNA usukuye ufite ibiranga bikurikira: umurongo uroroshye, A230 na A260 ni ingingo ebyiri, A300 yegereye 0, A260 / A280 = hafi 2.0, na A260 / A230 = hafi 2.0.Niba scan ya data itaboneka, igipimo cya A260 / A230 nacyo kigomba kugenwa, kuko iki gipimo cyunvikana cyane na carryover yimyanda yose igira ingaruka kumikorere ya enzymatique.Witondere umurongo ugizwe nigikoresho (0.1–0.5 kuri A260).

Hariho ibindi bintu bibiri byingirakamaro: igipimo kizaba hafi 0.3 munsi mugihe A260 / A280 apimye mumazi;mugihe igipimo cyapimwe muri 10 mM EDTA kiri hejuru ya 0.2 kurenza icyapimwe muri 1 mM EDTA.

Ibicuruzwa bifitanye isano:

Ubushinwa Bwuzuye RNA Yigunga Ibikoresho Byakozwe nuwabitanga |Foregene (foreivd.com)

Urukurikirane rwa RNA rutanga Abaguzi ninganda |Ubushinwa RNA bwitandukanya bwabakora (foreivd.com)

Urukurikirane rwa RNA - Foregene Co, Ltd. (foreivd.com)