Ngwa akụrụngwa ngwa ngwa ngwa ngwa RNA
Atụmatụ
■ Dị Mfe na ngwa ngwa: Mwepụta RNA site na ihe nlele osisi nwere ike mezue n'ime nkeji iri atọ.
Compat Ndakọrịta siri ike: Ngwa a adabaghị naanị maka mkpụrụ akwụkwọ stem na akwụkwọ, kamakwa maka polysaccharide/polyphenol bara ụba.
■ Nchekwa na obere nsí: ihe ndị na-egbu egbu dị ka β-mercaptoethanol, DTT, chloroform, phenol adịghị mkpa.
Ngwa
Itable Kwesịrị ekwesị maka ịrụ ọrụ mgbada, gụnyere RT-PCR, RT-qPCR, ngwakọ ngwakọ, Northern Blot, Dot Blot, nyocha PolyA, ntụgharị in vitro, nyocha nchedo RNase na mgbachi molekụla, wdg.
Enwere ike ịhazi ngwaahịa niile maka ODM/OEM. Maka nkọwa,biko pịa Ọrụ ahaziri (ODM/OEM)
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Mkpokọta RNA nke ihe nlele akwụkwọ 65 mg (akwụkwọ klova, akwụkwọ hibiscus, akwụkwọ ọka wit, akwụkwọ osikapa, akwụkwọ mkpụrụ ugli, akwụkwọ Prunus cerasifera, akwụkwọ fig, akwụkwọ ụbọchị, akwụkwọ Kerria japonica), ihe nlele fungi 150 mg (Lentinus edodes) na 200 A na-ewepụta ihe nlele petal mg (petals lily) site na iji RNA Easy Fast Plant Tissue Kit. Ihe nyocha Bioanalyzer 2100 nke RNA zuru ezu sitere na petals lily. |
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Agilent Bioanalyzer 2100 nsonaazụ nsonaazụ RNA sitere na petals lily. |
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RNA wepụtara site n'omume dị iche iche edepụtara na tebụl site na iji RNA Easy Fast Plant Tissue Kit. Olu olu: 50 μl; Oke mkpọtụ: 2 μl. A na -eme electrophoresis na 6 V/cm maka nkeji 15 na agarose 1%. |
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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