Ngwa RNA dị mkpokọta RNA

Maka mmịpụta RNA zuru oke nke ọma site na iji kọlụm centrifugal a na-ejikarị.

Ngwakọta RNA dị obere RNA na -ewepụta ụzọ mwepụ RNA ọhụrụ dabere na guanidinium isothiocyanate/phenol. Buffer RZ nke TIANGEN haziri nke ọma nwere ike wepu DNA genomic na protein site na sel ngwa ngwa yana rụọ ọrụ nke ọma, na -eme ka RNA enwetara dị ọcha ma kwụsie ike.
A na -eji ngwa a ikewapụta RNA na ọbara, sel anụmanụ, anụ ahụ na anụ ahụ. Kọlụm nke ọ bụla nwere ike ịhazi anụ ahụ 50-100 mg ma ọ bụ 5 × 106selụ n'otu oge ma nwee ike hazie ọtụtụ ọnụọgụ dị iche iche n'otu oge. Enwere ike mezue mmeghachi omume n'ime ihe na -erughị otu elekere, na mkpokọta RNA ewepụtara nwere mkpụrụ dị elu, ịdị ọcha ka mma, na -enweghị mmetọ DNA na protein, enwere ike iji ya mee nnwale dị iche iche dị n'okpuru.

Nwamba. Mba Nkwakọ ngwaahịa
4992858 50 ngụkọta ọnụ

Nkọwa ngwaahịa

Usoro ọrụ

Nlereanya Nnwale

Ajụjụ

Mkpado ngwaahịa

Atụmatụ

NA RNA ịdị ọcha dị njikere iji dị mma maka ngwa mgbada mgbada.
Applications Ngwa sara mbara: Enwere ike itinye RNA dị ọcha na nlele nnwale dị iche iche.
■ Enwere ike mezue nnwale ahụ n'ime elekere 1 site na iji arụmọrụ dị mfe.

Ngwa

A na-akpọ RT-PCR
■ Northern Blot, ntụpọ ntụpọ.
PCR ezigbo oge
Nyocha PolyA, ntụgharị in vitro, nyocha nchedo RNase, cloning molekul.

Enwere ike ịhazi ngwaahịa niile maka ODM/OEM. Maka nkọwa,biko pịa Ọrụ ahaziri (ODM/OEM)


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    Workflow

    Experimental Example Ihe: 20 mg anụ ahụ splin
    Usoro: Mkpokọta RNA nke anụ ahụ na -efe efe dịpụrụ adịpụ site na iji RNAsimple Total RNA Kit.
    Nsonaazụ: Biko lee foto agarose gel electrophoresis dị n'elu. Ejiri 2-4 μl nke 100 μl eluates kwa otu ụzọ. A na -eme electrophoresis na 6 V/cm maka nkeji 30 na agarose 1%.
    Ajụjụ: Mkpọchi kọlụm

    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.

    Ajụjụ: Mkpụrụ RNA dị ala

    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.

    Ajụjụ: mbibi RNA

    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.

    Ajụjụ: mmetọ DNA

    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.

    Ajụjụ: Etu esi ewepu RNase na ihe nnwale nnwale na enyo enyo?

    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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