rube isi na nkwekọrịta ahụ", na-agbaso ihe achọrọ n'ahịa, na-esonye na asọmpi ahịa site na ịdị mma ya dị ka ọ na-enye nkwado zuru oke na nkwado maka ndị ahịa ka ha bụrụ nnukwu mmeri.The achụ nke ụlọ ọrụ, bụ maa ndị ahịa' ụtọ maka China New Product Best Selling Viral DNA&Rna Extraction Kit, Anyị gbasaa azụmahịa anyị n'ime Germany, Turkey, Canada, USA , Indonesia, India, Nigeria, Brazil na ụfọdụ ndị ọzọ na mpaghara nke ụwa.Anyị na-agbasi mbọ ike ịbụ otu n'ime ndị na-ebubata ngwaahịa kacha mma n'ụwa niile.
rube isi na nkwekọrịta ahụ", na-agbaso ihe achọrọ n'ahịa, na-esonye na asọmpi ahịa site na ịdị mma ya dị ka ọ na-enye nkwado zuru oke na nkwado maka ndị ahịa ka ha bụrụ nnukwu mmeri.Na-achụ nke ụlọ ọrụ, bụ maa ndị ahịa' ụtọ makaNgwa Mwepụta Rna & DNA nke China na Mwepụta Nucleic Acid, Anyị na-atụ anya ịmepụta mmekọrịta bara uru na gị dabere na ngwaahịa na ngwọta anyị dị elu, ọnụ ahịa ezi uche na ọrụ kacha mma.Anyị na-atụ anya na ngwaahịa anyị ga-ewetara gị ahụmahụ dị ụtọ ma na-ebu mmetụta nke ịma mma.
Akwụkwọ ntuziaka:Ngwakọta mkpokọta RNA kewapụrụ ihe ọkụkụ gbakwunyere ntuziaka ntuziaka

rube isi na nkwekọrịta ahụ", na-agbaso ihe achọrọ n'ahịa, na-esonye na asọmpi ahịa site na ịdị mma ya dị ka ọ na-enye nkwado zuru oke na nkwado maka ndị ahịa ka ha bụrụ nnukwu mmeri.The achụ nke ụlọ ọrụ, bụ maa ndị ahịa' ụtọ maka China New Product Best Selling Viral DNA&Rna Extraction Kit, Anyị gbasaa azụmahịa anyị n'ime Germany, Turkey, Canada, USA , Indonesia, India, Nigeria, Brazil na ụfọdụ ndị ọzọ na mpaghara nke ụwa.Anyị na-agbasi mbọ ike ịbụ otu n'ime ndị na-ebubata ngwaahịa kacha mma n'ụwa niile.
China New Product China Rna&DNA Extraction Kit na Nucleic Acid mmịpụta, Anyị na-atụ anya guzosie ike ibe gị mmekọrịta dabere na anyị elu-edu ngwaahịa na ngwọta, ezi uche ahịa na kasị mma ọrụ.Anyị na-atụ anya na ngwaahịa anyị ga-ewetara gị ahụmahụ dị ụtọ ma na-ebu mmetụta nke ịma mma.

Akwụnyere kọlụm

Ka etinyechara kọlụm ahụ, mkpụrụ RNA na-ebelata ma ọ bụ ọ gaghị ekwe omume ime ka RNA dị ọcha, yana oke RNA enwetara dị ala.

Nyocha ihe kpatara ya:

1. Ntụle nkwụsịtụ adịghị nke ọma.

Njiji ihe atụ anaghị eme ka egbochie kọlụm DNA-CLEANING kpamkpam, ebe ọ na-emetụta mkpụrụ RNA na ịdị mma.Anyị na-akwado ngwa ngwa-egweri ọrụ na a zuru ezu mmiri mmiri nitrogen mgbe ị mebiri samples, Gbalịa na-echipịa sample cell mgbidi, cell akpụkpọ ahụ na ndị ọzọ na anụ ahụ.Maka nlele osisi nke polyol polysaccharides, anyị na-akwado ka ị jiri Plant Total RNA ISOLATION KIT PLUS.

2. Mgbe ejiri DNA-Cleaning Kọlụm na-amịpụta ihe nlere nke kewapụrụ ekewa, enwere ike ikuru mmiri na-agbajisị cell.

Sedimentị gbajiri sel nke ewepụrụ ga-ebute kọlụm RNA-ONLY nke a ga-egbochi mgbe arụ ọrụ mgbasa ozi RNA na-arụ (lee nzọụkwụ 6).Anyị na-akwado gị nke ọma mgbe ị na-amịpụta ihe a na-ahụ anya iji zere irighiri irighiri cell.

3. Sample mbụ ego dị ukwuu.

Ojiji nke oke nlere ga-ebute nkewa nlere ezughị ezu ma ọ bụ lysis cell ezughị ezu site na Buffer PSL1, na-ebute mgbochi nke kọlụm nchacha n'oge a na-asachapụ.Ngwakọta mkpokọta RNA kewapụrụ ihe ọkụkụ nke ọ bụla n'otu n'otu arụrụ arụ ọrụ bụ 50 mg.Maka ihe atụ osisi nke polyol polysaccharides, anyị na-akwado ka ị nwaa Plant Total RNA ISOLATION KIT PLUS.

4. Okpomọkụ nke centrifuge dị oke ala.

A na-eme ikewapụ RNA niile na usoro ịsacha na ụlọ okpomọkụ (20-25°C), ma e wezụga na mmiri mmiri nitrogen agbajiwo anụ ahụ sample. Okpomọkụ nke ụfọdụ centrifuges cryogenic dị ala karịa 20℃, nke nwere ike ibute ngọngọ nke kọlụm DNA-Cleaning na/ma ọ bụ naanị kọlụm RNA.Ọ bụrụ na nke a emee, debe okpomọkụ centrifuge na 20-25℃, najide n'aka na ngwakọta lysis na/ma ọ bụ ethanol gbakwunyere na-ekpo ọkụ na-ekpo ọkụ na 37°C.

Ọ nweghị RNA ewepụtara ma ọ bụ mkpụrụ RNA dị ala

A na-enwekarị ọtụtụ ihe na-emetụta arụmọrụ mgbake, dị ka: sample ọdịnaya RNA, usoro ọrụ, elution olu, wdg.

Nyocha nke ihe na-akpatakarị dị ka n'okpuru:

1.A na-eme centrifugation ice ma ọ bụ obere okpomọkụ (4 ° C) n'oge ọrụ ahụ.

Aro: Na-arụ ọrụ na ụlọ okpomọkụ (15-25°C) na usoro dum, emela ice bat na obere okpomọkụ centrifugation.

2. A na-emebi RNA ahụ n'ihi nchekwa na-ezighị ezi nke ihe nlele ma ọ bụ nchekwa ogologo oge nke ihe nlele ahụ.

Nkwanye: A ga-ekpochapụ ihe nlele ndị anakọtara ọhụrụ na nitrogen mmiri mmiri ngwa ngwa, wee chekwaa ya na -80 Celsius ruo ogologo oge, zere ịza oyi ugboro ugboro na thawing samples;ma ọ bụ gbanye ihe nlele ozugbo na ngwọta RNA stabilizer RNAlater (ihe nlele anụmanụ).

3.Enweghị oke nlele nkewa na lysis na-eduga na ngọngọ nke kọlụm dị ọcha.

Aro: Mgbe ị na-egweri anụ ahụ, biko hụ na anụ ahụ agbadala nke ọma, ma bufee ya ngwa ngwa na Buffer PSL1 akwadoro (kwenye na agbakwunyere oke β-ME ziri ezi, lee nzọụkwụ 1 nke usoro).

4.Agbakwunyere eluent na-ezighi ezi.

Aro: Gbaa mbọ hụ na ddH2O na-enweghị RNase na-adaba n'etiti akpụkpọ ahụ kọlụm dị ọcha.

5.Etinyeghị olu ziri ezi nke ethanol zuru oke na Buffer PSL2 ma ọ bụ Buffer PRW2.

Aro: Biko soro ntuziaka a, tinye nha ziri ezi nke ethanol zuru oke na Buffer PSL2 na Buffer PRW2 wee gwakọta nke ọma tupu ejiri ngwa ahụ.

6. Ọnụ ọgụgụ nke anụ ahụ adịghị mma.

Aro: Jiri 50 mg nke anụ ahụ kwa 500 μl nke Buffer PSL1.Iji anụ ahụ buru ibu ga-ebelata ọnụọgụ RNA ewepụtara yana ịdị ọcha nke RNA ga-esi na ya pụta ga-ebelatakwa.Anyị na-akwado ike na usoro nlele mbụ ekwesịghị gafere 50 mg kwa ọrụ mmịpụta RNA.

7.Na-ekwesịghị ekwesị elution olu ma ọ bụ ezughị ezu elution.

Aro: Oke eluent nke kọlụm dị ọcha bụ 50-200 μl;Ọ bụrụ na mmetụta elution adịghị afọ ojuju, a na-atụ aro ịgbatị oge n'ime ụlọ okpomọkụ mgbe agbakwunyere ddH2O na-enweghị ọkụ RNase, dị ka 5-10min.

8.The kọlụm dị ọcha nwere ethanol fọdụrụnụ mgbe ịsacha na BufferPRW2.

Aro: Ọ bụrụ na centrifuged tube efu maka 1 min ma a ka nwere ethanol fọdụrụ mgbe ịsachara na Buffer PRW2, ị nwere ike ịbawanye oge nke tube centrifugation efu na 2 min, ma ọ bụ tinye kọlụm dị ọcha na ụlọ okpomọkụ maka 5 min iji wepụ ethanol fọdụrụnụ kpamkpam.

9.A na-eji ngwa ahụ eme ihe na-ezighi ezi.

Aro: Maka ihe atụ osisi nke polysaccharides polyphenolic, iji ngwa ndị a na-ahụkarị dịka Plant Total RNA Isolation Kit nwere ike ọ gaghị enwe ike nweta ụdị RNA dị mma.Anyị na-akwado ka ị jiri Plant Total RNA IsolationKit Plus, nke emebere nke ọma maka nlele osisi polyphenolic polysaccharide.Ngwa emepụtara nke ọma maka ịwepụta RNA na ihe atụ osisi polyphenol na polysaccharide.

OD260/OD280 uru dị ala

RNA elution na ddH2O ma jiri ya mee ihe maka ọgụgụ spectrophotometer na-arụpụta ụkpụrụ OD260/OD280 dị ala.Anyị na-akwado iji 10 mM Tris-HCl, pH 7.5 (kama RNase-Free ddH2O iji elute RNA) iji nweta ụkpụrụ OD260/OD280 ziri ezi, lee "Ntụle RNA Concentration and Purification Assays" na ibe 19.

A na-emebi RNA dị ọcha

Ọdịmma RNA dị ọcha metụtara ihe ndị dị ka ichekwa nlele, mmetọ RNase, na nhịahụ.

Nyocha nke ihe na-akpatakarị:

A naghị echekwa ihe nlele 1.Tissue na oge mgbe nchịkọta.

Nkwanye: Ọ bụrụ na ejighị ihe nlele anụ ahụ n'oge nchịkọta, biko chekwaa ha na mmiri mmiri nitrogen na obere okpomọkụ ozugbo ma ọ bụ bufee ya na -80 Celsius maka nchekwa ogologo oge mgbe oyi na-ekpo ọkụ ngwa ngwa na nitrogen mmiri mmiri, ma ọ bụ gbanye ihe nlele ozugbo na ngwọta RNA stabilizer RNAlater (ntụle anụmanụ).Maka mmịpụta RNA, gbalịa iji nlele anụ ahụ anakọtara ọhụrụ.

2.Na-emegharị ugboro ugboro na thawing nke anụ ahụ n'omume.

Aro: Mgbe ị na-echekwa ihe nlele anụ ahụ, ọ kacha mma ịkpụ ha n'ime obere iberibe maka nchekwa, wee wepụta akụkụ ha mgbe ị na-eji ha iji zere mmebi nke RNA nke oyi na-eme ugboro ugboro na agbaze nke samples.

A na-ewebata 3.RNase n'ime ụlọ ọrụ ma ọ bụ na-eyighị uwe, mkpuchi, wdg.

Aro: A na-eme nnwale mmịpụta RNA kacha mma na arụ ọrụ RNA dị iche iche, a ga-ehichapụkwa tebụl ụlọ nyocha tupu nnwale ahụ, a ga-eyiri uwe aka na ihe mkpuchi nwere ike iyi n'oge nnwale ahụ iji zere mbibi RNA kpatara site na iwebata RNase ruo oke.

4.The reagent na-emerụ site RNase n'oge eji.

Aro: Dochie na usoro ọhụrụ nke ngwa mkpokọta RNA ihe ọkụkụ maka nnwale ndị metụtara ya.

5.The centrifuge tubes na pipette ndụmọdụ eji eme ihe RNA na-emetọ ya na RNase.

Aro: Gbaa mbọ hụ na tubes centrifuge, ndụmọdụ pipette, pipettes, wdg. ejiri na mmịpụta RNA bụ RNase-efu.

Akwụkwọ ntuziaka:

Ngwakọta mkpokọta RNA kewapụrụ ihe ọkụkụ gbakwunyere ntuziaka ntuziaka