TRNzol Universal Reagent
Atụmatụ
Flexibility Mgbanwe dị elu: oke ọhịa nke ịmalite nlele, dabara maka nnukwu mmịpụta nlele olu n'otu mmeghachi omume.
Yield Mkpụrụ dị elu: Usoro mmiri ozuzo na -eme ka mkpụrụ nke RNA dị na nlele.
Use Ojiji zuru oke: Kwesịrị ekwesị maka ọtụtụ nlele dị iche iche dị ka anụ na anụ ụlọ, mkpụrụ ndụ omenala, ọbara, mmiri ahụ, wdg.
Operation Ọrụ ngwa ngwa: enwere ike nweta DNA nke anụ ahụ n'ime elekere 1.
Nkọwapụta
Ụdị: Mmiri na -adabere
Ịlele: Nje virus, nje bacteria, ero, anụ, anụ ahụ, ihe ọkụkụ, sel na azụ mmiri.
Ebumnuche: RNA
Oge ọrụ: ~ 1 elekere
Ngwa: TRNzol Universal reagent na -ebelata mmetọ nke adịghị ọcha dịka DNA na protein n'ime mkpokọta RNA dị ọcha, enwere ike iji ya mee ihe maka nnwale ihe omimi dị iche iche dị ka Northern Blot, Dot Blot, nyocha PolyA, ntụgharị in vitro, nyocha nchedo RNase, owuwu ụlọ akwụkwọ cDNA. , RT-PCR, PCR ozugbo na usoro usoro dị elu.
Enwere ike ịhazi ngwaahịa niile maka ODM/OEM. Maka nkọwa,biko pịa Ọrụ ahaziri (ODM/OEM)
Usoro: 30 mg anụ ahụ imeju oke, akwụkwọ nri osikapa 100 mg anakọtara site na mmiri mmiri nitrogen na -egweri; 1 × 106HepG2 sel ndị a zụlitere na 700 μl Saccharomyces Cerevisiae medium (OD600 = 0.9) anakọtara site na centrifugation. 1 ml nke TRNzol Universal Reagent sitere na TIANGEN yana ngwaahịa ndị dị mkpa sitere na ndị na -ebubata L na T agbakwunyere na ntinye nke nlele ọ bụla wee wepụta RNA na -eso usoro iwu nke onye na -eweta ngwaahịa ọ bụla nyere. Olu elution bụ 80 μl, 50 μl, 30 μl na 30 μl maka nlele anọ ahụ n'otu n'otu. Ejiri 3 μl nke elu elu kwa otu ụzọ.
MIII: TIANGEN akara III;
A na -eme electrophoresis na 6 V/cm maka nkeji 30 na agarose 1%.
Nsonaazụ: TIANGEN TRNzol Universal Reagent nwere ike wepụta ịdị ọcha dị elu na ezi iguzosi ike n'ezi ihe RNA site na imeju oke, akwụkwọ osikapa, mkpụrụ ndụ emebere eme na ihe nlele yist, yana arụmọrụ dị elu. Ogo RNA dị ka ma ọ bụ dịtụ elu karịa nke ndị na -eweta ngwaahịa L na T.
A-1 Cell lysis ma ọ bụ homogenization ezughị
---- Belata ojiji nlele, mụbaa ọnụọgụ ihe nchekwa lysis, mụbaa homogenization na oge lysis.
A-2 Ego nlele buru oke ibu
---- Belata ọnụ ọgụgụ nlele ejiri ma ọ bụ mụbaa ọnụọgụ ihe nchekwa lysis.
A-1 lysis sel zuru oke ma ọ bụ homogenization
---- Belata ojiji nlele, mụbaa ọnụọgụ ihe nchekwa lysis, mụbaa homogenization na oge lysis.
A-2 Ego nlele buru oke ibu
---- Biko rụtụ aka na ikike nhazi kacha.
Apụghị A-3 RNA kpamkpam na kọlụm
---- Mgbe agbakwunyere mmiri RNase-Free, hapụ ya maka nkeji ole na ole tupu ịmịcha.
A-4 Ethanol dị na elu
---- Mgbe ịsachasịrị, centrifuge ọzọ wee wepụ ihe ịsa ahụ dịka o kwere mee.
A-5 ewepụghị usoro ọdịbendị kpamkpam
---- Mgbe ị na-anakọta mkpụrụ ndụ, biko gbaa mbọ hụ na ị wepụrụ usoro ọdịbendị ka enwere ike.
A-6 Mkpụrụ ndụ echekwara na RNAstore anaghị agbasasị nke ọma
---- njupụta ụlọ ahịa RNA karịrị nkezi ọdịbendị sel; yabụ ekwesịrị ịbawanye ike centrifugal. A na -atụ aro ka centrifuge na 3000x g.
A-7 Ọdịnaya RNA dị ala na ụba na nlele
---- Jiri nlele dị mma chọpụta ma ọ bụrụ na nlele ahụ kpatara obere mkpụrụ.
A-1 Ihe a abụghị nke ọhụrụ
---- Ekwesịrị idobe anụ ahụ ọhụrụ na nitrogen mmiri mmiri ozugbo ma ọ bụ tinye ya ozugbo na reagent RNAstore iji hụ na mmịpụta ahụ.
A-2 Ego nlele buru oke ibu
---- Belata ego nlele.
A-3 RNase mmetọn
---- Ọ bụ ezie na ihe nchekwa enyere na ngwa ahụ enweghị RNase, ọ dị mfe imetọ RNase n'oge usoro mwepụ yana ekwesịrị iji ya mee nke ọma.
A-4 mmetọ Electrophoresis
---- Dochie ihe nchekwa electrophoresis wee gbaa mbọ hụ na ihe ndị na-eri ihe na ihe nchekwa na-enweghị mmetọ RNase.
A-5 Oke oke maka electrophoresis
---- Belata ego nlele nlele, itinye nke ọma ọ bụla ekwesịghị ịfe 2 μg.
A-1 Ego nlele buru oke ibu
---- Belata ego nlele.
A-2 Ụfọdụ nlele nwere ọdịnaya DNA dị elu ma enwere ike iji DNase gwọọ ya.
Mee ọgwụgwọ DNase na-enweghị RNase na ntinye RNA enwetara, enwere ike iji RNA mee ihe maka nnwale ndị na-esote mgbe ọgwụgwọ gasịrị, ma ọ bụ ihe mkpocha RNA nwere ike mee ka ọ dị ọcha.
Maka iko iko, sie na 150 Celsius maka awa 4. Maka arịa plastik, mikpuru n'ime 0.5 M NaOH maka nkeji iri, wee jiri mmiri na-enweghị RNase sachaa ya nke ọma wee sterilaiz iji wepụ RNase kpamkpam. Ihe reagents ma ọ bụ azịza ejiri mee nnwale ahụ, ọkachasị mmiri, ga -abụrịrị enweghị RNase. Jiri mmiri na-enweghị RNase maka nkwadebe reagent niile (gbakwunye mmiri na karama iko dị ọcha, gbakwunye DEPC na mkpokọta ikpeazụ nke 0.1% (V/V), maa jijiji n'otu abalị na autoclave).
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