teerera chibvumirano”, zvinoenderana nezvinodiwa pamusika, anojoinha kubva mumakwikwi emusika nemhando yayo yakanaka zvakare sezvo ichipa rutsigiro rwakakwana uye rwakanyanya kune vatengi kuti vavarege vachikunda.Iko kutsvaga kwekambani, iko kunakidzwa kwevatengi kuChina Chigadzirwa Chitsva Chinotengeswa Viral DNA & Rna Extraction Kit, Takawedzera bhizinesi redu muGermany, Turkey, Canada, USA, Indonesia, India, Nigeria, Brazil uye mamwe matunhu epasi.Tiri kushanda nesimba kuti tive mumwe wevanonyanya kutengesa pasi rose.
teerera chibvumirano”, zvinoenderana nezvinodiwa pamusika, anojoinha kubva mumakwikwi emusika nemhando yayo yakanaka zvakare sezvo ichipa rutsigiro rwakakwana uye rwakanyanya kune vatengi kuti vavarege vachikunda.Iko kutsvaga kwekambani, ndiko kunakidzwa kwevatengiChina Rna & DNA Extraction Kit uye Nucleic Acid Kubvisa, Tiri kutarisira kumisa hukama hunobatsira pamwe newe zvichibva pane zvigadzirwa zvedu zvemhando yepamusoro uye mhinduro, mitengo inonzwisisika uye sevhisi yakanakisa.Tinovimba kuti zvigadzirwa zvedu zvichakuunzira chiitiko chinofadza uye kutakura kunzwa kwekunaka.
Mirayiridzo yezvinyorwa:Dyara Yese RNA Isolation Kit Plus Instruction Manual

teerera chibvumirano”, zvinoenderana nezvinodiwa pamusika, anojoinha kubva mumakwikwi emusika nemhando yayo yakanaka zvakare sezvo ichipa rutsigiro rwakakwana uye rwakanyanya kune vatengi kuti vavarege vachikunda.Iko kutsvaga kwekambani, iko kunakidzwa kwevatengi kuChina Chigadzirwa Chitsva Chinotengeswa Viral DNA & Rna Extraction Kit, Takawedzera bhizinesi redu muGermany, Turkey, Canada, USA, Indonesia, India, Nigeria, Brazil uye mamwe matunhu epasi.Tiri kushanda nesimba kuti tive mumwe wevanonyanya kutengesa pasi rose.
China Chigadzirwa Chitsva China Rna & DNA Extraction Kit uye Nucleic Acid Kubvisa, Tiri kutarisira kumisa hukama hunobatsira pamwe newe zvichibva pane zvigadzirwa zvedu zvemhando yepamusoro uye mhinduro, mitengo inonzwisisika uye sevhisi yakanakisa.Tinovimba kuti zvigadzirwa zvedu zvichakuunzira chiitiko chinofadza uye kutakura kunzwa kwekunaka.

Iyo column yakavharwa

Mushure mekunge koramu yavharwa, goho reRNA rinodzikiswa kana kutotadza kuchenesa iyo RNA, uye iyo yakawanikwa RNA misa yakaderera.

Kazhinji chikonzero chekuongorora:

1. Sample breaks haisi yakakwana.

Sample kuputsika hakukonzerese zvachose kuti DNA-CLEANING COLUMN ivharwe, uku ichikanganisa goho reRNA uye mhando.Isu tinokurudzira kukurumidza kukuya kushanda mune yakakwana mvura nitrogen pawakatyora masampuli, Edza kupwanya sampuli sero madziro, sero membrane uye mamwe matishu.Kune zvidyarwa zvemasampunzi e polyol polysaccharides, isu tinokurudzira kuti iwe ushandise Plant Yese RNA IsolATION KIT PLUS.

2. Kana ukasveta sample yakapatsanurwa supernatant neDNA-Cleaning Column, sero rinobvira rakatsemuka mvura inogona kufemerwa.

Iyo sero yakatsemuka sediments yakatorwa inokonzeresa iyo RNA-CHETE Column iyo ichavharika kana RNA adsorption oparesheni yaitwa (ona danho rechitanhatu).Isu tinokukurudzira nekuchenjerera paunosveta iyi supernatant kudzivirira maseru mamarara ari kuyamwa.

3. Sample yekutanga mari yakawandisa.

Kunyanya kushandiswa kwemuenzaniso kuchaita kuti isapera kupatsanurwa kwemuenzaniso kana kusakwana sero lysis neBuffer PSL1, zvichikonzera kuvharika kwekoramu yekuchenesa panguva yekucheneswa.Plant Yese RNA Isolation Kit Imwe neimwe yakanatswa inoshanda sample i50 mg.Kune zvidyarwa zvemasampunzi e polyol polysaccharides, isu tinokurudzira kuti uedze Kudyara Yese RNA IsolATION KIT PLUS.

4. Kupisa kwe centrifuge kwakanyanya kuderera.

Iyo yese RNA yekuzviparadzanisa uye yekucheneswa maitiro inoitwa pane yekudziya tembiricha (20-25°C), kunze kwekunge iyo sampuli tishu yakatyoka ne nitrogen yemvura. Tembiricha yemamwe cryogenic centrifuges yakaderera kupfuura 20.℃, izvo zvinogona kukonzera kuvharika kweDNA-Cleaning Column uye/kana RNA-Chete Column.Kana izvi zvikaitika, isa tembiricha ye centrifuge kusvika 20-25℃,uyeita shuwa kuti musanganiswa welysis uye/kana iyo ethanol-yakawedzerwa supernatant yakapiswa kusvika 37°C.

Hapana RNA yakabviswa kana RNA goho rakaderera

Pane kazhinji zvinhu zvakawanda zvinokanganisa kugona kudzoreredza, senge: sampuli RNA yemukati, nzira yekushanda, iyo elution vhoriyamu, nezvimwe.

Kuongororwa kwezvikonzero zvakajairika sezvinotevera:

1.Kushamba kwechando kana kutonhora kwakadzika (4 ° C) centrifugation yakaitwa panguva yekushanda.

Zano: Shanda patembiricha yekamuri (15-25°C) mukuita kwese, usaite ice bath uye yakaderera tembiricha centrifugation.

2.RNA yakasvibiswa nekuda kwekuchengetedzwa kusina kunaka kwemuenzaniso kana kuchengetedza kwenguva refu kwemuenzaniso.

Kurudziro: Zvichangobva kuunganidzwa samples dzinofanirwa kukurumidza kuomeswa nechando mumvura ine nitrogen, yozochengetwa pa -80°C kwenguva yakareba, kudzivisa kuomesa nechando uye kunyungudutswa kwemasampuli;kana kuti nekukasira kunyudza masampuli muRNA stabilizer RNAlater mhinduro (samples dzemhuka).

3.Kusakwana kupatsanurwa kwemuenzaniso uye lysis kunotungamirira kukuvhara kwekoromo yekuchenesa.

Mazano: Paunenge uchikuya mutsara, tapota chengetedza kuti mutsara wakakwana pasi, uye nekukurumidza uende nayo kuBuffer pre-prepared PSL1 (simbisa kuti chikamu chakarurama che β-ME chakawedzerwa, ona danho rekutanga rekuita).

4.The eluent yakawedzerwa zvisizvo.

Kurudziro: Ita shuwa kuti RNase-Yemahara ddH2O yakadonhedzwa pakati pechenese column membrane.

5.Iyo chaiyo vhoriyamu yeabsolute ethanol haina kuwedzerwa kuBuffer PSL2 kana Buffer PRW2.

Zvaungaita: Tapota tevera mirairo, wedzera huwandu chaihwo hweabsolute ethanol kuBuffer PSL2 neBuffer PRW2 uye sanganisa zvakanaka kititi isati yashandiswa.

6.Kuwanda kwemuenzaniso wenyama hakuna kufanira.

Zano: Shandisa 50 mg yetishu pa500 μl yeBuffer PSL1.Kushandisa nyama yakawandisa kunoderedza huwandu hweRNA yakabviswa uye kuchena kweiyo RNA inoguma kunodzikiswa zvakare.Isu tinokurudzira zvakasimba kuti yekutanga sampuli dosage haifanire kudarika 50 mg paRNA yekubvisa oparesheni.

7.Inappropriate elution volume kana kusakwana.

Zano: Vhoriyamu yakasarudzika yekoramu yekuchenesa ndeye 50-200 μl;kana iyo elution mhedzisiro isiri inogutsa, zvinokurudzirwa kuwedzera nguva pakamuri tembiricha mushure mekuwedzera preheated RNase-Mahara ddH2O, senge 5-10min.

8.Koramu yekuchenesa ine ethanol yakasara mushure mekugeza neBufferPRW2.

Zano: Kana iyo chubhu isina chinhu iine centrifuged ye1 min uye ichine ethanol yakasara mushure mekugezwa muBuffer PRW2, unogona kuwedzera nguva yechubhu isina chinhu centrifugation kusvika ku2 min, kana kuisa mbiru yekuchenesa pakupisa kwekamuri ye5 min kuti ubvise zvachose ethanol yakasara.

9.Kiti yakashandiswa zvisizvo.

Zano: Kune zvidyarwa zvepolyphenolic polysaccharides, uchishandisa zvakajairika kits sePlant Yese RNA Isolation Kit inogona kusakwanisa kuwana yakakodzera RNA samples.Isu tinokukurudzira kuti ushandise Plant Yese RNA IsolationKit Plus, iyo yakanyatsogadzirirwa polyphenolic polysaccharide chirimwa samples.Kiti yakagadziridzwa kuburitsa RNA kubva ku polyphenol uye polysaccharide chirimwa samples.

OD260/OD280 kukosha kwakaderera

RNA elution ine ddH2O uye inoshandiswa kuverenga spectrophotometer inoguma yakaderera OD260/OD280 kukosha.Tinokurudzira kushandisa 10 mM Tris-HCl, pH 7.5 (panzvimbo peRNase-Yemahara ddH2O kuti utore RNA) kuti uwane zvakati tsvikiti zveOD260/OD280, ona “RNA Concentration and Purification Assays” papeji 19.

Iyo RNA yakacheneswa yakaderedzwa

Hunhu hweRNA yakacheneswa hune hukama nezvinhu zvakaita sekuchengetedza sampuli, kusvibiswa kweRNase, uye kunyengera.

Kuongororwa kwezvikonzero zvinowanzoitika:

1.Tissue samples haana kuchengetwa munguva mushure mekuunganidza.

Kurudziro: Kana masampuli ematishu asina kushandiswa munguva mushure mekuunganidzwa, ndapota achengetedze mu nitrogen yemvura pa tembiricha yakaderera pakarepo kana kuti uendese kune -80 ° C kuitira kuchengetedza kwenguva refu mushure mekukurumidza kutonhora mu nitrogen yemvura, kana kunyudza nekukurumidza masampuli muRNA stabilizer RNAlater solution (mhuka samples ).Nekubvisa RNA, edza kushandisa masampula ematishu achangobva kuunganidzwa.

2.Kudzokororwa kwechando uye kunyunguduka kwemasampuli ematishu.

Zvaungaita: Paunenge uchichengeta matishu esampuli, zviri nani kuacheka kuita zvidimbu zvidiki kuti zvichengeteke, uye wotora chikamu chawo paunenge uchiashandisa kudzivirira kuparara kweRNA kunokonzerwa nekuomesa nechando kakawanda uye kunyunguduka kwemasamples acho.

3.RNase inounzwa mukamuri yekuvhiya kana kusapfekwa magirovhosi anoraswa, masiki, nezvimwe.

Zano: RNA kutorwa kwekuyedza kunoitwa zvakanyanya mumabasa akasiyana eRNA, uye tafura yemurabhoritari inofanirwa kucheneswa kusati kwaedzwa, uye magirovhosi nemasks anoraswa anofanira kupfekwa panguva yekuyedza kudzivirira kuparara kweRNA kunokonzerwa nekuunzwa kweRNase zvakanyanya.

4.Iyo reagent yakasvibiswa neRNase panguva yekushandiswa.

Zano: Tsiva nenhevedzano itsva yechirimwa yakazara RNA yekubvisa kits yezviyedzo zvinoenderana.

5.The centrifuge tubes uye pipette mazano anoshandiswa paRNA manipulation akasvibiswa neRNase.

Zvaungaita: Ita shuwa kuti machubhu ecentrifuge, matipi epipette, mapipetti, nezvimwewo zvinoshandiswa muRNA kudhirowa zvese zviri RNase-Mahara.

Mirayiridzo yezvinyorwa:

Dyara Yese RNA Isolation Kit Plus Instruction Manual