Izixhobo zethu ezixhotyiswe kakuhle kunye nolawulo olukumgangatho ophezulu kuwo onke amanqanaba okwenziwa kwemveliso kusenza siqinisekise ukoneliseka komthengi ngokupheleleyo kwiFactory ngokuthe ngqo iChina Techstar Nest Mgpure Nucleic Acid yokuTshala iKit Rna Isolation Kit, "Ukwenza iiMveliso kunye nezisombululo zoMgangatho Omkhulu" inokuba yinjongo yanaphakade yombutho wethu.Senza iinzame ezingagungqiyo zokuqonda injongo yokuba “Siza kuhlala sikwiNgqiqo ngamaxesha onke”.
Amaziko ethu axhotyiswe kakuhle kunye nolawulo lomgangatho olungileyo kuwo onke amanqanaba okwenziwa kusenza ukuba siqinisekise ukoneliseka komthengi ngokupheleleyoChina Nucleic Acid Reagent, DNA / Rna Extraction Kit, Sinikezela ngeenkonzo ezifanelekileyo, impendulo ekhawulezileyo, ukuhanjiswa kwangexesha, umgangatho obalaseleyo kunye nexabiso elingcono kubathengi bethu.Ukwaneliseka kunye nekhredithi elungileyo kuye wonke umthengi yinto ephambili yethu.Sijolisa kuzo zonke iinkcukacha zokucwangciswa kweodolo kubathengi bade bafumane iimveliso ezikhuselekileyo nezivakalayo ezinenkonzo yolungiselelo kunye neendleko zoqoqosho.Kuxhomekeke koku, iimveliso zethu zithengiswa kakuhle kakhulu kumazwe ase-Afrika, kuMbindi-Mpuma kunye nakuMzantsi-mpuma Asia.

Iinkcukacha

I-50 Preps, i-200 Preps Ikiti isebenzisa ikholomu ye-spin kunye nefomula ephuhliswe yi-Foregene, enokuthi ikhuphe ngokufanelekileyo i-RNA ephezulu kunye nomgangatho ophezulu we-RNA kwiintlobo ezahlukeneyo zezityalo ezine-polysaccharides okanye umxholo we-polyphenols.Inika ikholomu ye-DNA-Cleaning enokuthi isuse ngokulula i-DNA ye-genomic kwi-supernatant kunye ne-tissue lysate.Ikholamu ye-RNA kuphela inokubopha ngokufanelekileyo i-RNA.Ikhithi inokuqhuba inani elikhulu leesampuli ngexesha elinye.

Inkqubo yonke ayinayo i-RNase, ngoko ke i-RNA ecocekileyo ayiyi kuthotywa.I-Buffer PRW1 kunye ne-Buffer PRW2 inokuqinisekisa ukuba i-RNA efunyenweyo ayingcoliswanga yiprotheni, i-DNA, i-ion, kunye ne-organic compounds.

Amacandelo ekhithi

Iimpawu&izinto eziluncedo

■ Ukusebenza kwiqondo lobushushu begumbi (15-25℃) kuyo yonke inkqubo, ngaphandle kokuhlamba komkhenkce kunye nobushushu obuphantsi be-centrifugation.■ Gqibezela ikhithi ye-RNase-Free, akukho mfuneko yakukhathazeka malunga nokuthotywa kwe-RNA.■ Ngokukodwa ifanele ukucocwa kwe-RNA kwiisampuli zezityalo ze-polysaccharides kunye ne-polyphenols.∎ IKholam yokucoca i-DNA ibophelela ngokukodwa kwi-DNA, ukuze ikhithi ikwazi ukususa ungcoliseko lwe-genomic DNA ngaphandle kokongeza i-DNase.■ Isivuno esiphezulu se-RNA: IKholamu ye-RNA kuphela kunye nefomula ekhethekileyo inokucoca ngokufanelekileyo i-RNA.■ Isantya esikhawulezayo: kulula ukusebenza kwaye sinokugqitywa kwimizuzu engama-30.■ Ukhuseleko: akukho sixhobo sokwenza izinto eziphilayo esifunekayo.∎ Umgangatho ophezulu: Amaqhekeza e-RNA acociweyo anobunyulu obuphezulu, awanaprotheyini kunye nobunye ukungcola, kwaye anokumelana nemifuniselo eyahlukahlukeneyo esezantsi komlambo.

Iiparamitha zemveliso

■ Ukusetyenziswa komlambo osezantsi: i-cDNA synthesis ye-first-strand, i-RT-PCR, i-molecular cloning, i-Northern Blot, njl. njl. ■ Isampulu: Izihlunu zesityalo esitsha okanye esikhenkcezisiweyo se-polysaccharides kunye ne-polyphenols ■ Umthamo: 50mg izicubu zezityalo ■ Ubuninzi bomthamo wokubopha i-RNA kwikholamu yokucoca: 80 μg 20 ■ 50 Elution

Isicelo seKit

Ifanelekile ukutsalwa kunye nokucocwa kwe-RNA epheleleyo kwiisampuli zezicubu zezityalo ezitsha okanye ezikhenkcezisiweyo (ingakumbi izicubu zeqabunga lesityalo esitsha) kunye nomxholo ophezulu we-polysaccharide kunye ne-polyphenol.

Ukuhamba komsebenzi

Umzobo

Isityalo esipheleleyo se-RNA Isolation Kit Plus icutshungulwe i-50mg yamagqabi amatsha e-polysaccharides kunye ne-polyphenols, kunye ne-5% ecocekileyo ye-RNA yavavanywa yi-electrophoresis.1: Ibhanana 2: IGinkgo 3: Umqhaphu 4: Irharnati

Ugcino kunye nobomi beShelf

Le khithi ingagcinwa iinyanga ezingama-24 phantsi kweemeko ezomileyo kumaqondo obushushu begumbi (15-25℃);ukuba ifuna ukugcinwa ixesha elide, inokugcinwa kwi-2–8℃.I-Buffer PSL1 inokubekwa kwi-4℃ kwinyanga eyi-1 emva kokongeza i-β-mercaptoethanol (kuyacetyiswa ukuba yongezwe ngexesha elifanayo lovavanyo). Izixhobo zethu ezixhotyiswe kakuhle kunye nolawulo oluphezulu lomgangatho ophezulu kuzo zonke izigaba zokuvelisa kusenza ukuba siqinisekise ulwaneliseko lomthengi olupheleleyo lweFactory ngokuthe ngqo China Techstar Nest Mgpure Mgpure Mgpure I-Nest Mgpure I-Nest Mgpure Imveliso ye-Kit I-Nucleic Acid ye-Purification Kit Kit Kit. rge Quality” isenokuba yinjongo engunaphakade yentlangano yethu.Senza iinzame ezingagungqiyo zokuqonda injongo yokuba “Siza kuhlala sikwiNgqiqo ngamaxesha onke”.
Factory ngqoChina Nucleic Acid Reagent, DNA / Rna Extraction Kit, Sinikezela ngeenkonzo ezifanelekileyo, impendulo ekhawulezileyo, ukuhanjiswa kwangexesha, umgangatho obalaseleyo kunye nexabiso elingcono kubathengi bethu.Ukwaneliseka kunye nekhredithi elungileyo kuye wonke umthengi yinto ephambili yethu.Sijolisa kuzo zonke iinkcukacha zokucwangciswa kweodolo kubathengi bade bafumane iimveliso ezikhuselekileyo nezivakalayo ezinenkonzo yolungiselelo kunye neendleko zoqoqosho.Kuxhomekeke koku, iimveliso zethu zithengiswa kakuhle kakhulu kumazwe ase-Afrika, kuMbindi-Mpuma kunye nakuMzantsi-mpuma Asia.

Uluhlu lufakiwe

Emva kokuba ikholamu iplagwe, isivuno se-RNA sincitshisiwe okanye akunakwenzeka ukuhlambulula i-RNA, kwaye ubunzima obufunyenweyo be-RNA buphantsi.

Uhlalutyo lwesizathu esiqhelekileyo:

1. Uqhawulo lwesampulu alucokisekanga.

Ukuqhekeka kwesampuli akubangeli ngokupheleleyo i-DNA-CLEANING COLUMN ukuba ivalwe, ngelixa ichaphazela isivuno se-RNA kunye nomgangatho.Sincoma umsebenzi wokusila ngokukhawuleza kwi-nitrogen engamanzi eyaneleyo xa uqhekeza iisampuli, Zama ukutyumza udonga lweseli yesampuli, inwebu yeseli kunye nezinye iithishu.Kwiisampuli zezityalo ze-polyol polysaccharides, sincoma ukuba usebenzise i-Plant Total RNA IOLATION KIT PLUS.

2. Xa ufunxa isampulu eyohluliweyo ye-supernatant nge-DNA-Cleaning Column, i-cell fragmented precipitate enokwenzeka inokuphefumlelwa.

Iintlenga eziqhekekileyo zeseli ezithathiweyo ziya kubangela iKholamu ye-RNA KUPHELA eya kuvalelwa xa kusenziwa umsebenzi we-adsorption we-RNA (bona inyathelo lesi-6).Siyakucebisa ngononophelo xa ufunxa le supernatant ukunqanda ubugoxo beeseli ukuba bufunxe.

3. Isixa sokuqala sesampulu sininzi kakhulu.

Ukusetyenziswa kwesampulu ngokugqithisileyo kuya kubangela ukuqhekeka kwesampulu engaphelelanga okanye i-cell lysis engaphelelanga yi-Buffer PSL1, okukhokelela ekuvalelekeni kwekholamu yokucoca ngexesha lokuhlanjululwa.I-Plant Total RNA Isolation Kit Isampuli nganye yokusebenza ecocekileyo yi-50 mg.Kwiisampulu zezityalo ze-polysaccharides ze-polyol, sincoma ukuba uzame i-Plant Total RNA IOLATION KIT PLUS.

4. Ubushushu be-centrifuge buphantsi kakhulu.

Yonke i-RNA yodwa kunye nenkqubo yokucoca iqhutyelwa kwindawo yokushisa (20-25°C), ngaphandle kokuba isampulu ithishu yaphulwe yinitrogen engamanzi.Ubushushu bezinye iicentrifuge zecryogenic bungaphantsi kwama-20.℃, enokubangela ukuvaleka kweKholamu yokucoca i-DNA kunye/okanye i-RNA-Only Column.Ukuba oku kwenzeka, setha ubushushu be-centrifuge ukuya kuma-20-25℃, kwayeqinisekisa ukuba umxube we-lysis kunye/okanye i-ethanol-eyongeziweyo ye-supernatant yayifudunyezwe ibe ngama-37°C.

Akukho RNA ekhutshiweyo okanye isivuno se-RNA siphantsi

Ngokuqhelekileyo kukho izinto ezininzi ezichaphazela ukusebenza kakuhle kokubuyisela, njenge: umxholo we-RNA yesampuli, indlela yokusebenza, umthamo we-lution, njl.

Uhlalutyo lwezizathu eziqhelekileyo zingezantsi:

I-1.I-ice bath okanye ukushisa okuphantsi (4 ° C) i-centrifugation yenziwa ngexesha lokusebenza.

Icebiso: Isebenza kwiqondo lobushushu begumbi (15-25°C) kuyo yonke inkqubo, musa ukwenza i-ice bath kunye ne-centrifugation yobushushu obuphantsi.

I-2.I-RNA iye yachithwa ngenxa yokugcinwa okungafanelekanga kwesampuli okanye ukugcinwa kwexesha elide lesampuli.

Isindululo: Iisampulu ezisandula ukuqokelelwa kufuneka zikhe zikhenkcezwe ngokukhawuleza kwinitrogen engamanzi, emva koko zigcinwe ku -80°C ixesha elide, zithintele ukuphinda kukhenkceze kunye nokunyibilika kweisampulu;okanye ngokukhawuleza ucwilise iisampuli kwi-RNA stabilizer RNAlater isisombululo (iisampulu zezilwanyana).

I-3.Ukwahlulwa kwesampulu enganelanga kunye ne-lysis kukhokelela ekuvinjweni kwekholamu yokucoca.

Isiphakamiso: Xa ugaya i-tissue, nceda uqinisekise ukuba i-tissue iphantsi ngokwaneleyo, kwaye ngokukhawuleza uyidlulisele kwi-Buffer ye-PSL1 elungiselelwe ngaphambili (qinisekisa ukuba umlinganiselo ochanekileyo we-β-ME wongezwe, jonga inyathelo 1 lenkqubo).

4.I-eluent yongezwa ngokungalunganga.

Ingcebiso: Qinisekisa ukuba i-ddH2O ye-RNase-Free ithontsizelwe embindini we-membrane yekholamu yokucoca.

5.Umthamo ochanekileyo we-ethanol epheleleyo ayizange yongezwe kwi-Buffer PSL2 okanye i-Buffer PRW2.

Ingcebiso: Nceda ulandele imiyalelo, yongeza umthamo ochanekileyo we-ethanol epheleleyo kwi-Buffer PSL2 kunye ne-Buffer PRW2 kwaye udibanise kakuhle phambi kokuba ikhithi isetyenziswe.

6.Ubungakanani besampulu yethishu ayifanelekanga.

Ingcebiso: Sebenzisa i-50 mg yethishu nge-500 μl ye-Buffer PSL1.Ukusebenzisa izicubu ezininzi kakhulu kuya kunciphisa inani le-RNA ekhutshiweyo kwaye ukucoceka kwe-RNA okubangelwayo kuya kuncitshiswa.Sincoma kakhulu ukuba idosi yesampulu yokuqala ayifanele idlule i-50 mg ngomsebenzi wokutsalwa kwe-RNA.

7.Ivolume ye-elution engafanelekanga okanye i-elution engaphelelanga.

Isiphakamiso: Umthamo ocacileyo wekholamu yokucoca ngu-50-200 μl;ukuba i-elution effect ayinelisi, kuyacetyiswa ukuba kwandiswe ixesha kwiqondo lobushushu legumbi emva kokongeza i-ddH2O ye-RNase-Free preheated, efana ne-5-10min.

I-8.Ikholamu yokucoca inentsalela ye-ethanol emva kokuhlamba nge-BufferPRW2.

Isiphakamiso: Ukuba ityhubhu engenanto i-centrifuged ye-1 min kwaye kusekho i-ethanol eseleyo emva kokuhlamba kwi-Buffer PRW2, unokwandisa ixesha le-tube engenanto ye-centrifugation ukuya kwi-2 min, okanye ubeke ikholamu yokucoca kwindawo yokushisa ye-5 min ukususa ngokupheleleyo i-ethanol eseleyo.

9.Ikhithi isetyenziswe ngokungachanekanga.

Isiphakamiso: Kwiisampuli zezityalo ze-polyphenolic polysaccharides, ukusebenzisa iikiti eziqhelekileyo ezifana ne-Plant Total RNA Isolation Kit ayinakukwazi ukufumana iisampulu ezifanelekileyo ze-RNA.Sincoma ukuba usebenzise iPlant Total RNA IsolationKit Plus, eyilelwe ngokukodwa iisampuli zezityalo ze-polyphenolic polysaccharide.Ikhithi ephuhliswe ngokukodwa ukukhupha i-RNA kwi-polyphenol kunye neesampuli zezityalo ze-polysaccharide.

Ixabiso le-OD260/OD280 liphantsi

I-RNA elution ene-ddH2O kwaye isetyenziselwa ukufundwa kwe-spectrophotometer iphumela kumaxabiso aphantsi e-OD260/OD280.Sincoma ukusebenzisa i-10 mM Tris-HCl, i-pH 7.5 (kunokuba i-RNase-Free ddH2O yokukhupha i-RNA) ukufumana amaxabiso achanekileyo e-OD260/OD280, bona “i-RNA Concentration and Purification Assays” kwiphepha le-19.

I-RNA ehlanjululweyo ithotywa

Umgangatho we-RNA ecocekileyo inxulumene nezinto ezifana nokugcinwa kwesampulu, ukungcoliseka kwe-RNase, kunye nokukhwabanisa.

Uhlalutyo lwezizathu eziqhelekileyo:

I-1.Iisampulu ze-tissue azizange zigcinwe ngexesha emva kokuqokelela.

Isincomo: Ukuba iisampulu zezicubu azisetyenziswanga ngexesha emva kokuqokelela, nceda uzigcine kwi-nitrogen engamanzi kwiqondo lokushisa eliphantsi ngokukhawuleza okanye uzidlulisele kwi--80 ° C yokugcina ixesha elide emva kokuqhwala ngokukhawuleza kwi-nitrogen engamanzi, okanye ngokukhawuleza ucwilise iisampuli kwi-RNA stabilizer RNAlater isisombululo (iisampulu zezilwanyana).Ukukhutshwa kwe-RNA, zama ukusebenzisa iisampulu zeethishu ezisanda kuqokelelwa.

2.Ukukhenkceza okuphindaphindiweyo kunye nokunyibilika kweesampuli zethishu.

Icebiso: Xa ugcina iisampulu zethishu, kungcono ukuzisika zibe ziingceba ezincinci ukuze zigcinwe, kwaye ukhuphe inxalenye yazo xa uzisebenzisa ukuthintela ukonakaliswa kweRNA okubangelwa kukukhenkcezwa okuphindaphindiweyo nokunyibilika kweisampulu.

3.I-RNase yaziswa kwigumbi lokusebenza okanye inganxitywanga iiglavu ezilahlwayo, iimaski, njl.

Ingcebiso: Imifuniselo yokutsalwa kwe-RNA yenziwa ngcono kwimisebenzi eyahlukeneyo ye-RNA, kwaye itafile yelabhoratri kufuneka icocwe phambi kovavanyo, kwaye iiglavu ezilahlwayo kunye nemaski kufuneka zinxitywe ngexesha lovavanyo ukunqanda ukuthotywa kwe-RNA okubangelwa kukwaziswa kwe-RNase ngowona mgangatho mkhulu.

I-4.I-reagent ingcolisekile yi-RNase ngexesha lokusetyenziswa.

Ingcebiso: Faka endaweni yoluhlu olutsha lwezixhobo ze-RNA zokutsalwa kwezityalo zizonke zemifuniselo enxulumeneyo.

I-5.Iibhubhu ze-centrifuge kunye neengcebiso ze-pipette ezisetyenziselwa ukuguqulwa kwe-RNA zingcolile nge-RNase.

Ingcebiso: Qinisekisa ukuba iityhubhu ze-centrifuge, iingcebiso ze-pipette, ii-pipettes, njl njl. ezisetyenziswa kwi-RNA extraction zonke zi-RNase-Free.