Ngwa RNAprep Pure Plant Plus
Atụmatụ
Get Ebumnuche: A haziri ya nke ọma maka nlele osisi nke siri ike iwepụ, dị ka polysaccharides na osisi bara ọgaranya polyphenolics. A na -emezi usoro ahụ nke ọma, nsonaazụ ya bụ ntụkwasị obi.
Removal Mwepụ nke gDNA nke ọma: A na-enye DNase I nke na-arụ ọrụ nke ọma maka iwepụ gDNA ngwa ngwa na kọlụm.
■ Ọ dị mfe yana ngwa ngwa: nnwale mmịpụta RNA nwere ike mezue n'ime elekere 1.
Nchekwa na obere nsí: ọ dịghị ihe na -emepụta ihe na -egbu egbu dị ka phenol na chloroform dị mkpa.
Ngwa
Enwere ike iji ngwa a maka nnwale bayoloji dị iche iche dịka RT-PCR, PCR oge, Northern Blot, Dot Blot, nyocha PolyA, ntụgharị in vitro, nyocha nchedo RNase, na mkpuchi, wdg.
Enwere ike ịhazi ngwaahịa niile maka ODM/OEM. Maka nkọwa,biko pịa Ọrụ ahaziri (ODM/OEM)
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Enwetara mkpokọta RNA site na anụ ahụ nke 100 mg, anyụ, apụl na ube, tubers ụtọ nduku na nduku, akwụkwọ owu, rose, alfalfa, osikapa na agịga pine ọcha n'otu n'otu site na iji RNAprep Pure Plant Plus Kit. Ejiri 4-6 μl nke 30 μl eluates kwa otu ụzọ. Nna -ukwu: TIANGEN akara III; A na -eme electrophoresis na 6 V/cm maka nkeji 30 na agarose 1%. Nsonaazụ: RNAprep Pure Plant Plus Kit nwere ike wepụta ịdị ọcha dị elu, mkpụrụ dị elu na ezigbo iguzosi ike n'ezi ihe zuru oke RNA site n'omume osisi polysaccharides & polyphenolics. |
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).
Ngwaahịa ngwaahịa
N'IHI gịnị ka ịhọrọ anyị
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