Ọbara Mwepu RNA
Nkọwa ngwa:
Nwamba. Mba.RE-04011/04013
Maka mkpokọta RNA dị ọcha site na ọbara dum 104 ≤Mkpụrụ ndụ ọbara ọcha ≤ 107
Wepụ ngwa ngwa ma sachaa ọbara RNA na mkpụrụ ndụ ọbara ọcha.
- Ọ dịghị mkpa ichegbu onwe gị maka mmebi RNA.Ngwa niile bụ RNase-efu
-Mfe-arụ ọrụ niile na-agwụ agwụ na ụlọ okpomọkụ
- Ngwa ngwa-arụ ọrụ nwere ike mechaa n'ime nkeji iri abụọ
-Mkpụrụ RNA dị elu: naanị kọlụm RNA na usoro pụrụ iche nwere ike sachaa RNA nke ọma
-Nchekwa — ọ dịghị organic reagent eji
-Ike nhazi nhazi nke ukwuu-ihe ruru 200μl nwere ike ịhazi oge ọ bụla.
Ọdịmma dị elu - RNA emechara dị ọcha nke ukwuu, enweghị protein na adịghị ọcha ndị ọzọ, ọ nwekwara ike zute ngwa nnwale dị iche iche nke ala.
Nkọwa
Ngwa ahụ na-anabata kọlụm ogho na usoro nke ụlọ ọrụ anyị mepụtara, nke nwere ike wepụta RNA dị ọcha na ịdị elu nke ọma site na ọbara niile na-egbochi ọrịa.Ngwa ahụ na-enye lysate ọbara ọbara uhie (Buffer RCL), nke nwere ike imebi mkpụrụ ndụ ọbara uhie ngwa ngwa na nke ọma ma na-ejigide sel ọbara ọcha.Kọlụlụ DNA- Cleaning nke ọma nwere ike kewaa supernatant na cell lysates na mgbasa ozi ma wepụ DNA genomic.Arụ ọrụ dị mfe ma na-azọpụta oge;Kọlụm naanị RNA nwere ike jikọta RNA nke ọma, yana iji usoro pụrụ iche, ọ nwere ike hazie ọnụọgụ nlele n'otu oge.
Usoro niile RNase-Free na-eme ka RNA ewepụtara ghara imebi;Buffer RW1 na Buffer RW2 sistemu ịsacha ihe na-eme ka RNA enwetara enweghị protein, DNA, ion na mmetọ ngwakọta organic.
Ọdịnaya ngwa
| Ngwakọta Mkpokọta Ọbara RNA | ||
| Ngwa mejupụtara | RE-04011 | RE-04013 |
| ugboro 50 | ugboro 200 | |
| Ihe nchekwa RCL (10×) | 52.5 ml | 210ml |
| Ihe nchekwa BRL1* | 30ml | 120ml |
| Ihe nchekwa BRL2 | 18ml | 66ml |
| Ihe nchekwa RW1* | 25ml | 100ml |
| Ihe nchekwa RW2 | 24ml | 96 ml |
| ddH na-efu RNase2 O | 10ml | 40ml |
| RNA-naanị kọlụm | 50 tent | 200 setịpụ |
| Kọlụm na-ehicha DNA | 50 tent | 200 setịpụ |
| akwụkwọ ntuziaka | 1 oyiri | 1 oyiri |
Atụmatụ & uru
- Ọ dịghị mkpa ichegbu onwe gị maka mmebi RNA.Ngwa ahụ niile enweghị RNase.
-Mfe-arụ ọrụ niile na-agwụ agwụ na ụlọ okpomọkụ.
- Ngwa ngwa-arụ ọrụ nwere ike mechaa n'ime nkeji iri abụọ.
-Mkpụrụ RNA dị elu: naanị kọlụm RNA na usoro pụrụ iche nwere ike sachaa RNA nke ọma.
-Nchekwa — ọ dịghị organic reagent eji.
-Ike nhazi nhazi nke ukwuu-ihe ruru 200μl nwere ike ịhazi oge ọ bụla.
Ọdịmma dị elu - RNA emechara dị ọcha nke ukwuu, enweghị protein na adịghị ọcha ndị ọzọ, ọ nwekwara ike zute ngwa nnwale dị iche iche nke ala.
Ngwa ngwa
Ngwa ngwa:
Ọ dabara maka mmịpụta na ịdị ọcha nke mkpokọta RNA site na ọbara dum anụ mammali.
Usoro ọrụ
Ọnọdụ nchekwa
Ekwesịrị ịchekwa ihe nchekwa RCL (10 ×) na 2-8 ℃;Enwere ike ịchekwa ihe ndị ọzọ nke ngwa ahụ na ụlọ okpomọkụ (15-25 ℃) n'okpuru ọnọdụ akọrọ, enwere ike ịchekwa ya maka ọnwa 12.Enwere ike ịchekwa ihe nchekwa BRL1 na 4 ℃ maka ọnwa 1 ka ịgbakwunye β-mercaptoethanol (nhọrọ) .
Rịba ama: Ọ bụrụ na echekwara ya na obere okpomọkụ, ngwọta na-adịkarị mfe mmiri ozuzo.Jide n'aka na ị ga-etinye ihe ngwọta na kit na ụlọ okpomọkụ maka oge tupu ejiri ya.Ọ bụrụ na ọ dị mkpa, kpoo ya na mmiri ịsa mmiri 37 Celsius C maka nkeji 10 iji gbazee mkpọda ahụ, ma gwakọta nke ọma tupu eji ya.
Ntuziaka maka nyocha nsogbu
The following is an analysis of the problems that might be encountered in the extraction of viral RNA. We wish it would be helpful to your experiment. In addition, for other experimental or technical problems other than operating instructions and problem analysis, we have dedicated technical support to help you. Contact us if you need at : 028-83361257or E-mail：Tech@foregene.com。
Enweghị RNA enweghị ike ịwepụta ma ọ bụ mkpụrụ nke nucleic acid dị ala
A na-enwekarị ọtụtụ ihe na-emetụta arụmọrụ mgbake, dị ka: sample ọdịnaya RNA, usoro ọrụ, elution volume, wdg.
Nyocha nke ihe na-akpatakarị:
1.Ice bath ma ọ bụ obere okpomọkụ (4 Celsius C) centrifugation n'oge ọrụ.
Aro: Arụ ọrụ ụlọ okpomọkụ (15-25 Celsius C), ọ dịghị mgbe ịsa ahụ na ice na obere okpomọkụ centrifuge.
2. Nchekwa ihe nlere na-ezighi ezi ma ọ bụ nchekwa nchekwa maka ogologo oge.
Aro: Chekwaa nlele na -80 Celsius ma ọ bụ ifriizi na mmiri mmiri nitrogen, ma zere iji friza eme ihe ugboro ugboro;nwaa iji ihe nlele anakọtara ọhụrụ maka mmịpụta RNA.
3.Ezughị ezu sample lysis
Nkwanye: Biko hụ na agwakọtala ihe nlele na ihe ngwọta na-arụ ọrụ (Linear Acrylamide) ma tinye ya maka nkeji 10 n'ime ụlọ okpomọkụ (15-25 Celsius C).
4.Agbakwunyere eluent na-ezighi ezi
Nkwanye: Gbaa mbọ hụ na agbakwunyere RNase-Free ddH2O n'etiti akpụkpọ ahụ nke kọlụm nchacha.
5.Ezigbo olu nke anhydrous ethanol na Buffer viRW2
Aro: Biko soro ntuziaka a, tinye nha kwesịrị ekwesị nke ethanol anhydrous na Buffer viRW2 wee gwakọta ha nke ọma tupu i jiri ngwa ahụ.
6.Ezigbo sample ojiji.
Aro: 200µl nke nlele kwa 500μl nke ihe nchekwa viRL.Oke sample oke ga-ebute mbelata mmịpụta RNA.
7.Ezigbo elution olu ma ọ bụ ezughị ezu elution.
Aro: Oke eluent nke kọlụm nchacha bụ 30-50μl;Ọ bụrụ na mmetụta elution adịghị afọ ojuju, a na-atụ aro ka ịtinye ddH na-enweghị ọkụ ọkụ nke RNase.2O ma gbatịa oge idowe na ụlọ okpomọkụ, dị ka 5-10min
8.Purification kọlụm nwere ethanol residue mgbe rinsing na Buffer viRW2.
Aro: Ọ bụrụ na ethanol ka dịgidere ka emechara ya na Buffer viRW2 na centrifugation tube efu maka nkeji 2, enwere ike ịhapụ kọlụm dị ọcha na ụlọ okpomọkụ maka 5min ka centrifugation na-enweghị tube ga-ewepụ kpamkpam ethanol fọdụrụnụ.
Mmebi nke ụmụ irighiri ihe RNA dị ọcha
Ọdịmma RNA emechara dị ọcha metụtara ihe ndị dị ka nchekwa nlele, mmetọ RNase, na ọrụ.
Nyocha nke ihe na-akpatakarị:
1. A naghị echekwa ihe ndị a na-anakọta n'oge.
Aro: Ọ bụrụ na ejighị ihe nlele ahụ n'oge nchịkọta, biko chekwaa ya na -80 ℃ ma ọ bụ nitrogen mmiri mmiri ozugbo.Maka mmịpụta nke ụmụ irighiri ihe RNA, gbalịa iji ihe nlele anakọtara ọhụrụ mgbe ọ bụla enwere ike.
2.Achịkọtara samples bụ kefriza na thawing ugboro ugboro.
Aro: Zenarị oyi na agbaze ugboro ugboro (ọ dịghị ihe karịrị otu ugboro) n'oge nchịkọta na nchekwa, ma ọ bụghị ya, mkpụrụ nke nucleic acid ga-ebelata.
E webatara 3.RNase n'ime ụlọ ịsa ahụ ma ọ bụ enweghị uwe, ihe mkpuchi, wdg.
Aro: Mwepụta nke nnwale ụmụ irighiri ihe RNA kacha mma n'ime ụlọ ọrụ RNA dị iche, a na-ehichakwa tebụl nnwale tupu nnwale ahụ.Yiri gloves na masks enwere ike iwepụ n'oge nnwale ahụ ka ịzenarị mmebi RNA nke iwebata RNase kpatara.
4.The reagent na-emerụ site RNase n'oge ojiji.
Aro: Dochie na Viral RNA Isolation Kit ọhụrụ maka nnwale ndị metụtara ya.
5.The RNase mmetọ nke centrifuge tubes, pipette ndụmọdụ, wdg Aro: Jide n'aka na centrifuge tubes, pipette ndụmọdụ, na pipettes niile bụ RNase-free.
Ụmụ irighiri ihe RNA emechara emetụta nnwale ndị dị n'okpuru ala
Ngwakọta RNA ndị a sachara site na kọlụm dị ọcha ga-emetụta nnwale ndị dị n'okpuru ma ọ bụrụ na enwere ion nnu ma ọ bụ protein dị ukwuu, dị ka: ntụgharị ntụgharị, Northern Blot, wdg.
1.There fọdụrụ ion nnu na ụmụ irighiri RNA eluted.
Nkwanye: Gbaa mbọ hụ na agbakwunyere oke nke ethanol anhydrous na Buffer viRW2, wee sachaa kọlụm nke ịdị ọcha ugboro abụọ dịka ọsọ centrifugation ziri ezi na ntuziaka ọrụ; Ọ bụrụ na ion nnu ka fọdụrụ, ị nwere ike itinye Buffer viRW2 na kọlụm nchacha, wee hapụ ya na ụlọ okpomọkụ maka 5min.Mgbe ahụ mee centrifugation iji wepụ mmetọ ion nnu ruo n'ókè kasị ukwuu
2.There fọdụrụ ethanol na eluted RNA ụmụ irighiri
Aro: ozugbo na-akwado na Buffer viRW2 sachara kọlụm nchacha, mee centrifugation tube efu dịka ọsọ centrifugal na ntuziaka ọrụ.Ọ bụrụ na ethanol ka fọdụrụ, enwere ike ịhapụ ya maka nkeji 5 n'ime ụlọ okpomọkụ mgbe centrifugation na-adịghị na tube ga-ewepụ ethanol fọdụrụnụ ruo n'ókè kasị ukwuu.
Akwụkwọ ntuziaka:
Akwụkwọ ntuziaka ngwa mwepu RNA dịpụrụ adịpụ
