Sterilization yepipette matipi uye EP machubhu, nezvimwe.
1. Gadzirira 0.1% (chiuru) DEPC (yakanyanya muchetura zvinhu) nemvura yakasvibiswa, ishandise zvakanyatsonaka muhutsi hwehutsi, uye uichengetedze pa4 ° C kure nechiedza;
DEPC mvura imvura yakachena inobatwa neDEPC uye yakasvibiswa nekupisa kwakanyanya uye kudzvanywa.Yakaedzwa kuve isina RNase, DNase uye proteinase.
2. Isa iyo pipette tip uye EP tube mu 0.1% DEPC, uye chengetedza kuti pipette tip uye EP tube yakazadzwa ne 0.1% DEP.
3. Dzivirira kubva kuchiedza, rega umire, husiku (12-24h)
4. Bhokisi rine tip uye EP tube haifaniri kuiswa muDEPC.Mushure mekubvisa zvakanyanyisa mvura yeDEPC mumuromo kana EP chubhu, itakure uye uiputire.
5. 121 madhigirii Celsius, 30min
6. 180 madhigirii celsius, yakaoma kwemaawa akati wandei (angangoita maawa matatu)
Cherechedza: a.Pfeka latex magurovhosi uye masiki paunenge uchibata DEPC!b, kana pasina DEPC sterilization, 130 ℃, 90min autoclave (marabhoritari mazhinji akakwira tembiricha sterilization kaviri)
RNA kutorwa kwekufunga
Zviitiko zviviri zvikuru zvetishu RNA yekuzviparadzanisa nevamwe kutadza
RNA kushatisa uye zvisaririra zvetsvina mumatishu,maererano nekuora, ngatitangei kutarisa kuti sei RNA yakatorwa kubva mumasero akagadzirwa isiri nyore kuderedzwa.Iripo RNA yekubvisa reagents ese ane zvikamu zvinokurumidza kurambidza RNase.Wedzera lysate kumasero akagadzirwa, uye ingoisanganisa, masero ose anogona kunyatsosanganiswa ne lysate, uye masero akazara lysed.Mushure mokunge masero ari lysed, zvinoshandiswa zvinoshandiswa mu lysate pakarepo zvinodzivisa intracellular RNase, saka RNA inoramba yakasimba.Izvi zvinoreva, nokuti masero akagadzirwa ari nyore uye anonyatsobatanidzwa ne lysate, RNA yavo haisi nyore kuderedzwa;kune rumwe rutivi, iyo RNA munyama inoderedzwa nyore nyore nekuti masero ari munyama haasi nyore kukurumidza kubata lysate.nekuda kwekusangana kwakakwana.Saka,tichifunga kuti pane nzira yekushandura matishu kuita sero rimwe chete uchidzivirira kuita kweRNA, dambudziko rekuora rinogona kugadziriswa zvachose.
Liquid nitrogen milling ndiyo inonyanya kushanda nzira yakadai.Nekudaro, iyo yemvura nitrogen milling nzira inonetsa, kunyanya kana nhamba yemasampuli yakakura.Izvi zvakakonzera chinhu chinotevera chakanakisa: iyo homogenizer.Thehomogenizernzira haifungi mubvunzo wekuti basa reRNase rinodzivirirwa sei masero asati asangana ne lysate, asi anonyengetera kuti chiyero chekuvhiringidza kwenyama kukurumidza kudarika chiyero icho intracellular RNase inosvibisa RN.
Mhedzisiro yemagetsi homogenizer iri nani,uye mhedzisiro yegirazi homogenizer yakashata, asi kazhinji, iyo homogenizer nzira haigone kudzivirira iyo degradation phenomenon.Nokudaro, kana kubudiswa kwakasvibiswa, homogenizer yemagetsi yepakutanga inofanira kushandiswa pakukuya ne nitrogen yemvura;iyo yepakutanga girazi homogenizer inofanira kuchinjwa kuva homogenizer yemagetsi kana yakananga yakagayiwa ne nitrogen yemvura.Dambudziko rinenge 100% rinogoneka.gadzirisa.
Dambudziko rekusachena rinokanganisa kuyedza kunotevera rine zvikonzero zvakasiyana-siyana pane kudzikisira, uye mhinduro dzakasiyana.Mukupedzisa,kana paine kushatisa kana kusvibiswa kwakasara munyama, nzira yekubvisa / reagent yechinhu chakananga chekuyedza inofanira kugadziridzwa.Haufanire kushandisa masamples ako akakosha pakuita optimization: unogona kutenga mhuka diki sehove/huku kubva pamusika, tora chikamu chinowirirana chezvinhu zvekubvisa RNA, uye chimwe chikamu chekubvisa mapuroteni - kugaya nemuromo, dumbu uye ura.
Iyo RNA yakanangwa yeRNA yakabviswa inoshandiswa kune dzakasiyana-siyana yekutevera miedzo, uye yayo yemhando zvinodiwa zvakasiyana
cDNA raibhurari kuvaka inoda RNA kutendeseka pasina masara e enzyme reaction inhibitors;Kumaodzanyemba kunoda yakakwirira RNA kutendeseka uye yakaderera zvinodiwa kune enzyme reaction inhibitors masara;RT-PCR haidi yakawandisa RNA kutendeseka,asi inodzivisa kuita kwe enzyme.Zvasara zvinodikanwa zvakasimba.Iyo inopinza inosarudza kubuda;pese apo chinangwa chiri chekuwana yakanyanya kuchena RNA, zvinodhura vanhu nemari.
Kuunganidza/Kuchengeta MaSamples
Zvinhu Zvinokanganisa Kudzikira Mushure memuenzaniso wasiya muviri mhenyu / kana yepakutanga nharaunda yekukura, iwo endogenous enzymes mumuenzaniso anotanga kusvibisa RNA,uye iyo degradation rate ine hukama nezviri mukati endogenous enzymes uye tembiricha.Nenzira, kune nzira mbiri chete dzekudzivisa zvachose endogenous enzyme basa: wedzera lysate pakarepo uye homogenize zvakakwana uye nokukurumidza;cheka kuita zvidimbu zvidiki uye pakarepo omesa mu nitrogen yemvura.Nzira mbiri idzi dzinoda kushanda nokukurumidza.Iyo yekupedzisira yakakodzera kune ese masampuli, nepo yekutanga inokodzera chete matishu ane yakaderera zvemukati maseru uye endogenous enzymes uye nyore kuita homogenize.Kunyanya, nyama yekudyara, chiropa, thymus, pancreas, spleen, uropi, mafuta, mhasuru tishu, nezvimwe zvinonyanya kuomeswa nechando nenitrogen yemvura isati yaenderera.
Kuparadzaniswa uye homogenization yemasampuli
Zvinhu Zvinokanganisa Kudzikira uye Goho Sample kupatsanurwa ndikokuitira homogenization yakakwana, iyo ndeyekuburitswa uye kuzere kweRNA.Masero anogona kuve akananga homogenized pasina kuputswa.Matishu anogona kuve homogenized chete mushure mekutyoka.Mbiriso uye mabhakitiriya zvinoda kutyorwa nema enzymes anoenderana asati agona kuenzaniswa.Matishu ane yakaderera endogenous enzyme content uye nyore homogenization inogona kupwanyika uye homogenized panguva imwe mu lysate ne homogenizer;chirimwa nyama, chiropa, thymus, pancreas, spleen, uropi, mafuta, mhasuru tishu uye mamwe masampuli, Angave akakwira endogenous enzymes kana kuti haaite homogenized nyore.saka kukanganisa kwenyama uye homogenization kunofanira kuitwa zvakasiyana.Iyo yakavimbika uye inobereka nzira yekuparadzanisa ndeyekugaya ne nitrogen yemvura, uye nzira yakavimbika yehomogenization ndeye kushandiswa kwemagetsi homogenizer.Chinyorwa chakakosha pamusoro pekugaya nenitrogen yemvura: sampuli haifanire kunyungudutswa panguva yese yekugaya, sezvo endogenous ma enzymes anogona kushanda kana akaomeswa nechando.
Sarudzo ye lysate
Kukanganisa kurerukira kwekushanda uye zvikonzero zvezvakasara zvekupedzisira tsvina Iyo inowanzoshandiswa lysis mhinduro inogona kupotsa inhibisa basa reRNase.Nokudaro, pfungwa inokosha yekusarudza sarudzo ye lysis ndeyokufunga pamwe chete nenzira yekuchenesa.Pane chinhu chimwe chete:masampuli ane yakakwira endogenous enzyme content anokurudzirwa kushandisa lysate ine phenol kuwedzera kugona kumisa endogenous enzymes.
Sarudzo yekuchenesa nzira
Zvinhu zvinokanganisa zvakasara endogenous tsvina, kudhirowa kumhanya Kune yakachena samples seseru, mhedzisiro inogutsa inogona kuwanikwa neinenge chero nzira yekuchenesa iripo.Asi kune mamwe akawanda masampuli, kunyanya ayo ane huwandu hwakanyanya hwetsvina senge zvidyarwa, chiropa, mabhakitiriya, nezvimwe, zvakakosha kusarudza nzira yekucheneswa yakakodzera.Iyo column centrifugal yekuchenesa nzira ine kukurumidza kuburitsa kumhanya uye inogona kunyatso bvisa tsvina inokanganisa inotevera enzymatic reaction yeRNA, asi inodhura (Forgene inogona kupa makiti anodhura, zvimwe zvakawanda tinya.here);uchishandisa zvehupfumi uye zvemhando yepamusoro nzira dzekuchenesa, senge LiCl kunaya, inogona zvakare kuwana zvinogutsa mhedzisiro, asi iyo nguva yekuvhiya yakareba..
"Kurangwa Kutatu uye Kutariswa Sere" kweRNA Kubvisa
Chirango 1:Kugumisa kusvibiswa kweexogenous enzymes.
Cherechedzo 1:Kunyatsopfeka masiki uye magurovhosi.
Cherechedzo 2:Iwo machubhu ecentrifuge, Misoro yeZano, matanda epipette, matangi e electrophoresis, uye mabhenji ekuedza akabatanidzwa mukuyedza anofanirwa kuraswa zvachose.
Cherechedzo 3:Iwo ma reagents / zvigadziriso zvinosanganisirwa mukuyedza, kunyanya mvura, inofanira kunge isina RNase-isina.
Chirango 2:Vhara basa re endogenous enzymes
Cherechedzo 4:Sarudza yakakodzera homogenization nzira.
Cherechedza 5:Sarudza lysate yakakodzera.
Cherechedza 6:Dzora huwandu hwekutanga hwemuenzaniso.
Chirango 3:Rondedzera chinangwa chako chekuwedzera
Cherechedza 7:Iine chero lysate system inoswedera kune yakanyanya kutanga huwandu hwesample, iyo yekuwedzera yekubudirira mwero inodonha zvakanyanya.
Cherechedza 8:Iyo chete chiyero chehupfumi chekubudirira RNA kudhirowa budiriro mune zvinotevera zviedzo, kwete goho.
Pamusoro 10 Zvitubu zveRNase Kusvibiswa
1. Zvigunwe ndizvo zvinotanga kuunza maenzayimu ekunze, saka magirovhosi anofanira kupfekwa nekutsiviwa nguva nenguva.Mukuwedzera, masks anofanirwawo kupfekwa, nekuti kufema zvakare kwakakosha sosi yema enzymes.Imwe bhenefiti yekupfeka girovhosi mask ndeye kuchengetedza muyedzeri.
2. Mazano epipette, centrifuge tubes, pipettes - RNase haigoni kushandiswa nekuita sterilization chete, saka mazano epipette uye centrifuge tubes inofanira kurapwa neDEPC, kunyange kana yakanyorwa seDEPC yakabatwa.Zvakanakisisa kushandisa pipette yakakosha-chinangwa, kuipukuta ne 75% doro rekotoni bhora risati rashandiswa, kunyanya tsvimbo;kuwedzera, iva nechokwadi kuti usashandise musoro wekubvisa.
3. Mvura/buffer inofanira kunge isina kusvibiswa kweRNase.
4. Kanenge tafura yekuedzwa inofanira kucheneswa ne 75% doro donje mabhora.
5.Endogenous RNase Matishu ese ane endogenous enzymes, saka kutonhora nekukurumidza kwematishu ane mvura nitrogen ndiyo nzira yakanakisa yekudzikisa kuderera.Iyo yemvura nitrogen yekuchengetedza / nzira yekukuya haina kunetsa, asi ndiyo yega nzira yematishu ane huwandu hwakanyanya hweendogenous enzymes.
6. RNA samples RNA extraction products inogona kunge iine zvisaririra zveRNase kusvibiswa.
7. Plasmid extraction Plasmid extraction inowanzoshandisa Rnase kusvibisa RNA, uye Rnase yakasara inofanira kugayiwa neProteinase K uye yakatorwa nePCI.
8. RNA kuchengetedza Kunyangwe ikachengetwa patembiricha yakaderera, tsvaga huwandu hweRNAse huchakonzeresa kuparara kweRNA.Mhinduro yakanakisisa yekuchengetedzwa kwenguva refu kweRNA ndeyekumisa munyu / doro, nokuti doro rinodzivisa zvose enzymatic basa pakudzika kwakadzika.
9. Apo cations (Ca, Mg) ine ions idzi, kupisa pa80C kwemaminitsi mashanu kuchaita kuti RNA iparadzwe, saka kana RNA inoda kupisa, sarudzo yekuchengetedza inoda kuva ne chelating agent (1mM Sodium Citrate, pH 6.4).
10. Maenzymes anoshandiswa mukuedza kunotevera anogona kunge akasvibiswa neRNase.
10 Mazano eRNA Kubvisa
1: Kurumidza kudzivirira RNase chiitiko.Samples inokurumidza kuomeswa mushure mekuunganidzwa, uye RNase inomiswa nekukurumidza kushanda panguva yelysis.
2: Sarudza nzira yakakodzera yekubvisa yetishu ine yakakwira ribozyme content, uye adipose tishu zviri nani kushandisa nzira ine phenol.
3: Kufanotaura kunaka kunoda Northern, cDNA raibhurari kuvakwa kunoda kutendeseka kwepamusoro, uye RT-PCR neRPA (Ribonuclease protection assay) haidi kutendeseka kwepamusoro.RT-PCR inoda kuchena kwakanyanya (enzyme inhibitor residues).
4: Kunyatsoita homogenization ndiyo kiyi yekuvandudza goho uye kuderedza kuderera.
5: Tarisa kuvimbika kweRNA electrophoresis kuonekwa, 28S: 18S = 2: 1 chiratidzo chakakwana, 1: 1 inogamuchirwawo kune dzakawanda zvidzidzo.
6: Kubviswa kweDNA yeRT-PCR, array analysis Zvakanakisisa kushandisa Dnase I kubvisa DNA.
7: Deredza kusvibiswa kwema enzyme exogenous - ma enzymes haagone kutengwa kunze.
8: Paunenge uchiisa pasi-concentration nucleic acid, co-precipitation reagent inofanira kuwedzerwa.Asi kudzivirira iyo co-precipitant ine enzymes uye DNA kusvibiswa.
9: Nyatsonyungudutsa iyo RNA, kana zvichidikanwa, kupisa pa65C kwemaminitsi mashanu.
nzira yekuchengetedza yakakodzera
Inogona kuchengetwa pa -20C kwenguva pfupi, uye pa -80C kwenguva yakareba.Danho rekutanga mukuvandudza goho reRNA nderekuziva kuti iyo RNA yemukati yemasampuli akasiyana inosiyana zvakanyanya.Kuwanda kwepamusoro (2-4ug / mg) zvakadai sechiropa, pancreas, mwoyo, huwandu hwepakati (0.05-2ug / mg) hwakadai sehuropi, embryo, itsvo, mapapu, thymus, ovary, low abundance (<0.05ug / mg) mg) zvakadai sedundira, bone, mafuta.
1: Lyse maseru kuburitsa RN - kana RNA ikasaburitswa, goho richaderedzwa.Magetsi homogenization inoshanda zvirinani pane dzimwe nzira dzekuita homogenization, asi ingangoda kusanganiswa nedzimwe nzira, senge liquid nitrogen mashing, enzymatic digestion (Lysozyme/Lyticase)
2: Optimization yenzira yekubvisa.Matambudziko makuru ane phenol-based nzira haana kukwana stratification uye chikamu RNA kurasikirwa (iyo supernatant haigone kubviswa zvachose).Kusakwana stratification kunokonzerwa nepamusoro nucleic acid uye mapuroteni emukati, ayo anogona kugadziriswa nekuwedzera huwandu hwe lysate inoshandiswa kana kuderedza huwandu hwesampuli.Nhanho yekubvisa chloroform yakawedzerwa kune adipose tissue.RNA kurasikirwa kunogona kuderedzwa nekudzosera-kupomba kana nekubvisa iyo organic layer inoteverwa necentrifugation.Dambudziko guru nekoromo centrifugation-yakavakirwa nzira ndeye yakawandisa sampuli.
Classic Extraction Tips
1. Phenol kucheneswa: Wedzera huwandu hwakaenzana hwe1: 1 Phenol / Chloroform uye funganisa nesimba kwemaminitsi 1-2.Centrifuge nekumhanya kukuru kwemaminetsi maviri.Nyatsobvisa mashura (80-90%).Usambofa wasvika pachikamu chepakati.Iyo yakaenzana vhoriyamu yemhinduro mhinduro inogona kuwedzerwa kuPhenol/Chloroform uye iyo supernatant yakabviswa.Iwo maviri ma supernatant anogona kusanganiswa pamwechete kuitira kunayisa kwe nucleic acid kuti goho rive nani.Usanyanya kupfava paunenge uchisanganisa, uye usaedze kubvisa mashura ese.
2. Kugeza ne 70-80% ethanol: Panguva yekugeza, nucleic acid inofanira kumiswa kuitira kuti munyu wasara wagezwa.Panguva imwecheteyo, pakarepo mushure mekudurura ethanol, centrifuge pahupamhi hwepamusoro kwemasekonzi mashomanana, uye wozobvisa ethanol yakasara nepipette.Dissolve mushure mekumira panzvimbo yekushisa kwema 5-10 maminitsi.
11. Kutorwa kwemasangano akakosha
1. Fibrous tissue: Kiyi yeRNA kutorwa kubva kune fibrous tissue yakadai semwoyo / skeletal muscle ndeyekuvhiringidza zvachose tishu.Aya matishu ane low cell density, saka huwandu hweRNA pahuremu hweyuniti huremu hwematishu hwakaderera, uye zviri nani kushandisa yakawanda yekutanga huwandu sezvinobvira.Iva nechokwadi chekukuya nyama yacho zvakakwana pasi pemamiriro echando.
2. Mishonga ine mapuroteni akakwirira / mafuta ehuwandu: huropi / yemiriwo yemafuta inokwana.Mushure mePCI kubviswa, iyo supernatant ine chena floccules.Iyo supernatant inofanira kubviswa zvakare ne chloroform.
3. Mishonga ine high nucleic acid / ribozyme content: spleen / thymus ine nucleic acid yakawanda uye ribozyme content.Kukuya matishu pasi pemamiriro echando anoteverwa nekukurumidza homogenization kunogona kuita ribozymes.Zvisinei, kana iyo lysate yakanyanyisa viscous (nekuda kwepamusoro nucleic acid content), iyo PCI yakatorwa haizokwanisi stratify zvinobudirira;kuwedzera mamwe lysate kunogona kugadzirisa nyaya iyi.Multiple PCI zvinyorwa zvinogona kubvisa mamwe masara eDNA.Kana mvura yakachena ikaumbwa pakarepo mushure mekuwedzera doro, inoratidza kusvibiswa kweDNA.Kubvisa zvakare ne acidic PCI mushure mekuparadzwa kunogona kubvisa kusvibiswa kweDNA.
4. Mushonga wemiti: Miti yemiti yakaoma zvikuru kupfuura mhuka yemhuka.Kazhinji, zvirimwa zvinogadzikwa pasi pemvura yenitrogen mamiriro, saka kuderedzwa kweRNA nema endogenous enzymes hakuna kujairika.Kana dambudziko rekusvibisa rikasagadziriswa, rinenge rakakonzerwa nekusachena kuri mumuenzaniso.Izvo zvisina kuchena zviri mumiti yakawanda zvinotungamira kune zvakasara, uye chikonzero chezvisaririra kazhinji nekuti izvi zvinosvibisa zvine zvimwe zvakafanana neRNA: iwe unonaya uye ini ndinonaya, uye iwe unoshambadza uye ini adsorb.Aya maitiro anoona kuti iwo akasimba kwazvo enzyme inhibitors.
Parizvino, kushambadzira RNA kuburitsa reagents kunogona kuchinjirwa kune angangoita ese mhuka tishu ane zvigadziriso zvidiki, asi kune mashoma ekutengesa eRNA anotora reagents anogona kunge akakodzera kune mazhinji ezvirimwa tishu.Neraki, Foregene inogona kupa yakakoshachirimwa RNA kuburitsa kits, tinePlant Yese RNA Isolation kit, Dyara Yese RNA Isolation kit Plus.Iyo yekupedzisira yakanyatsogadzirirwa zvirimwa zvine yakakwira polysaccharide uye polyphenol zvemukati.Kune RNA kudhirowa, mhinduro kubva kune vashandisi veLab yakanyanya kunaka.
12. Mhedzisiro yesaizi yechando uye kunyunguduka Sampu yechando inogona kunge yakakura, uye inoda kutemwa isati yashandiswa kuburitsa RNA.Mienzaniso inowanzonyunguduka (zvichida zvishoma) panguva yekucheka.Sampuli dzakaomeswa dzinogona kuyerwa kusati kwabviswa RNA, uye kunyungudika kuchaitika panguva iyi.Dzimwe nguva, kunyungudutswa kwemuenzaniso kunoitikawo panguva yemvura nitrogen milling process;kana sampuli yechando inowedzerwa zvakananga kune lysate pasina mvura nitrogen milling, uye kunyunguduka kuchaitika zvechokwadi pamberi pehomogenization yakakwana.Ongororo dzakaratidza kuti nyama yakaomeswa nechando inonyanya kukanganisa RNA panguva yekunyunguduka pane matishu matsva.Chingangove chikonzero: Iyo yechando-yanyungudika inovhiringa zvimiro mukati mesero, zvichiita kuti zvive nyore kuti endogenous enzymes ibatane zvakananga neRNA.
13. Kutonga kwehutano hweRNA Kazhinji, electrophoresis inoshandiswa kutonga kuvimbika kweRNA, uye A260 / A280 inoshandiswa kutonga kuchena kweRNA.Muchirevo, RNA yakasimba ine chiyero che 28S: 18S = 2.7: 1, uye data yakawanda inosimbisa chiyero che 28S: 18S = 2: 1.Ichokwadi ndechokuti inenge isina imwe yeRNA yakatorwa kubva kune dzimwe sampuli kunze kwemasero iri muchiyero che2: 1 (izvi zvakawanikwa uchishandisa Agilent Bioanalyzer).
Mhedzisiro ye electrophoresis yeRNA inobatwa nezvinhu zvakawanda, kusanganisira chimiro chechipiri, electrophoresis mamiriro, sampuli mutoro, dhigirii yekuzara neEB, etc. Shandisa native electrophoresis kuti uone RNA uye shandisa DNA Marker sekutonga.Kana iyo 28S pa 2kb uye 18S pa 0.9kb yakajeka, uye 28S: 18S > 1, kuvimbika kunogona kusangana nezvinodiwa zvezviedzo zvakawanda zvinotevera.
A260/A280 chiratidzo chakakonzera nyonganiso yakawanda.Chokutanga pane zvose, zvakakosha kujekesa zvinoreva chepakutanga chechiratidzo ichi che nucleic acids: yakachena RNA, iyo A260/280 = inenge 2.0.Yakachena RNA ndiyo 'chikonzero' uye A260/A280 = 2 ndiyo 'mhedzisiro'.Iye zvino munhu wese ari kushandisa A260 / A280 se 'chikonzero', achifunga kuti "kana A260 / A280 = 2, saka RNA yakachena", izvo zvinotungamirira mukuvhiringidzika.
Kana iwe uchifarira, iwe unogona kuwedzera reagent shoma iyo inowanzoshandiswa mukuchera, yakadai se phenol, guanidine isothiocyanate, PEG, nezvimwewo, kune yako RNA sampuli, uye wozoyera chiyero cheA260 / A280.Chokwadi ndechekuti mazhinji emareagents anoshandiswa kuburitsa RNA, pamwe nekusachena kwakawanda mumuenzaniso, anonwisa kutenderedza A260 neA280, inokanganisa A260/A280.
Iyo inonyanya kudzidzisa nzira parizvino ndeyekutarisa RNA samples mune iyo 200-300 nm renji.Iko kutenderera kweRNA yakachena ine zvinotevera maitiro: iyo curve yakatsetseka, A230 uye A260 maviri inflection mapoinzi, A300 iri pedyo ne0, A260/A280 = yakatenderedza 2.0, uye A260/A230 = yakatenderedza 2.0.Kana scan data isipo, chiyero cheA260/A230 chinofanirwawo kutariswa, sezvo reshiyo iyi inonyanya kunzwisiswa nekutakurwa kwetsvina yese inokanganisa maitiro enzymatic.Funga nezve mutsara weiyo mudziyo (0.1-0.5 yeA260).
Pane zvimwe zviviri zvinobatsira zviitiko: reshiyo ichave inenge 0.3 yakaderera apo A260 / A280 inopimwa mumvura;nepo reshiyo yakayerwa mu10 mM EDTA ingangoita 0.2 yakakwirira pane yakayerwa mu1 mm EDTA.
Zvigadzirwa zvinoenderana:
China Plant Yese RNA Isolation Kit Mugadziri uye Mutengesi |Foregene (foreivd.com)
RNA isolation series - Foregene Co., Ltd. (foreivd.com)
Nguva yekutumira: Jul-15-2022
