Xidhmada Unugga Dhirta Degdegga ah ee RNA Easy
Astaamaha
■ Fudud oo degdeg ah: Wadarta soo saaridda RNA ee muunadaha dhirta waxaa lagu dhammayn karaa 30 daqiiqo gudahood.
Compat La-qabsasho xoog leh: Qalabkani kuma habboona shay-baarka caleemaha iyo caleemaha caadiga ah, laakiin sidoo kale saamiyada hodanka ah ee polysaccharide/polyphenol.
■ Sunta nabdoon oo hooseysa: Walxaha sunta ah sida β-mercaptoethanol, DTT, chloroform, phenol looma baahna.
Codsiyada
Itable Ku habboon qalliinka hoose, oo ay ku jiraan RT-PCR, RT-qPCR, isku-darka isku-darka, Northern Blot, Dot Blot, baaritaanka PolyA, tarjumaadda fiitamiinka, falanqaynta ilaalinta RNase iyo xirmidda moodeelka, iwm.
Dhammaan alaabooyinka waxaa loo habeyn karaa ODM/OEM. Faahfaahinta,fadlan guji Adeegga La -habeeyey (ODM/OEM)
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Wadarta RNA ee shaybaarka caleenta 65 mg (caleemo dhego, caleemo hibiscus, caleemo qamadi, caleemo bariis, caleemo midho ugli, Prunus cerasifera caleemo, caleemo berde, caleemo maalmeed, caleemo japonica Kerria), 150 mg sambal fungus (Lentinus edodes) iyo 200 muunadaha tufaaxa mg (tufaaxa-maalin-ubax) ayaa la soo saaray iyadoo la adeegsanayo RNA Easy Fast Plant Tissue Kit. |
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Agilent Bioanalyzer 2100 natiijada falanqaynta wadarta RNA oo ka timid tufaaxa Day-ubaxa. |
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RNA waxaa laga soo saaray shaybaarro kala duwan oo ku qoran miiska iyadoo la adeegsanayo RNA Easy Plant Tissue Kit. Mugga digniinta: 50 μl; Mugga rarka: 2 μl. Qalabka korontada ayaa lagu sameeyay 6 V/cm muddo 15 daqiiqo ah 1% agarose. |
A-1 Lysis cell ama homogenization kuma filna
---- Yaree isticmaalka muunadda, kordhi xaddiga kaydinta lysis, kordhiso isugeynta iyo waqtiga lysis.
A-2 Tirada muunaddu aad bay u badan tahay
---- Yaree cadadka muunada la isticmaalay ama la kordhiyo tirada kaydinta lysis.
A-1 Lysis unug oo aan ku filnayn ama isugeyn
---- Yaree isticmaalka muunadda, kordhi xaddiga kaydinta lysis, kordhiso isugeynta iyo waqtiga lysis.
A-2 Tirada muunaddu aad bay u badan tahay
---- Fadlan tixraac awoodda wax-qabadka ugu badan.
A-3 RNA gebi ahaanba lagama soo qaadin tiirka
---- Kadib markaad ku darto biyaha RNase-Free, ka tag dhowr daqiiqo ka hor intaadan ku shubin.
A-4 Ethanol ee ugu sarreeya
---- Ka dib markii aad biyo raacisay, mar kale centrifuge ka saar bakhaarka dhaqidda intii suurtogal ah.
A-5 Dhaqanka dhexdhexaadka ah gebi ahaanba lagama saaro
---- Markaad uruurinayso unugyo, fadlan hubso inaad ka saarto dhexdhexaadinta dhaqanka intii suurtogal ah.
A-6 Unugyada lagu kaydiyo RNAstore si fiican looma darin
---- Cufnaanta RNAstore-ku waxay ka weyn tahay celceliska dhaqanka unugyada; sidaas darteed waa in la kordhiyaa ciidanka dhexe. Waxaa lagu talinayaa in centrifuge at 3000x g.
A-7 Wax yar oo RNA ka kooban iyo tiro badan oo muunadda ah
---- Isticmaal muunad togan si aad u go'aamiso in dhalidda hooseysa ay keentay sambalku.
A-1 Maaddadu ma aha mid cusub
---- Unugyada cusub waa in lagu kaydiyo nitrogen dareere ah isla markiiba ama isla markiiba lagu rido reagent RNAstore si loo hubiyo saamaynta soo saaridda.
A-2 Tirada muunaddu aad bay u badan tahay
---- Yaree cadadka saamiga.
A-3 RNase wasakhaysann
---- Inkastoo kaydka ku jira xirmada uusan ku jirin RNase, way fududahay in la sumoobo RNase inta lagu jiro habka soo saaridda waana in si taxaddar leh loo maareeyaa.
A-4 Nadiifinta Electrophoresis
---- Beddel keydka korontada oo hubso in alaabta wax-cunaysa iyo Kaydinta Loading ay ka madax bannaan yihiin faddaraynta RNase.
A-5 Rarid aad u badan oo loogu talagalay electrophoresis
---- Hoos u dhig xaddiga rarista muunadda, ceel kasta rarantiisu waa inaysan dhaafin 2 μg.
A-1 Tirada muunaddu aad bay u badan tahay
---- Yaree cadadka saamiga.
B
-U samee daaweynta DNase-ka-dhaafka ah ee DNase xalka RNA ee la helay, RNA-na waxaa si toos ah loogu adeegsan karaa tijaabooyinka xiga daaweynta kadib, ama waxaa lagu sii nadiifin karaa xirmooyinka is-nadiifinta RNA.
Muraayadaha dhalooyinka, waxaa lagu dubay 150 ° C 4 h. Weelasha balaastigga ah, oo lagu dhex milay 0.5 M NaOH muddo 10 daqiiqo ah, ka dibna si fiican loogu maydhay biyo aan RNase lahayn ka dibna jeermis si looga saaro RNase. Soo -saarayaasha ama xalka loo adeegsaday tijaabada, gaar ahaan biyaha, waa inay ka madax bannaan yihiin RNase. U isticmaal biyo aan RNase lahayn dhammaan diyaarinta reagent (ku dar biyo dhalo galaas nadiif ah, ku dar DEPC xaddiga ugu dambeeya ee 0.1% (V/V), rux habeen iyo autoclave).
Qaybaha alaabta
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