Jumlar Shuka Kayan Keɓewar RNA Jimlar RNA Purificaiton Kit don Shuka Ƙananan a cikin Polysaccharides da Polyphenols
Ƙayyadaddun bayanai
Shirye-shirye 50, Shirye-shiryen 200
Kit ɗin yana amfani da ginshiƙan juzu'i da dabara wanda Foregene ya haɓaka, wanda zai iya fitar da inganci mai inganci da ingantaccen jimillar RNA daga kyallen tsire-tsire masu ƙarancin polysaccharides da abun ciki na polyphenols.Don samfuran tsire-tsire masu manyan polysaccharides ko abun ciki na polyphenols, ana ba da shawarar yin amfani da Plant Total RNA Isolation Plus Kit don samun ingantacciyar sakamakon hakar RNA.Kit ɗin yana ba da ginshiƙin Tsabtace DNA wanda zai iya cire DNA a sauƙaƙe daga lysate mai girma da nama.Rukunin RNA-kawai zai iya ɗaure RNA daidai yadda ya kamata.Kit ɗin na iya aiwatar da babban adadin samfurori a lokaci guda.
Duk tsarin bai ƙunshi RNase ba, don haka tsarkakewar RNA ba zai ƙasƙanta ba.Buffer PRW1 da Buffer PRW2 na iya tabbatar da cewa RNA da aka samu ba a gurɓata su ta hanyar furotin, DNA, ions, da mahadi na halitta ba.
Abubuwan Kit
| Buffer PSL1, Buffer PS, Buffer PSL2 |
| Buffer PRW1, Buffer PRW2 |
| RNase-Free ddH2O, Rukunin Tsabtace DNA |
| Rukunin RNA-Kawai |
| Umarni |
Fasaloli & fa'idodi
■ Aiki a dakin da zafin jiki (15-25 ℃) a ko'ina cikin tsari, ba tare da wankan kankara da ƙananan zafin jiki ba.
n Cikakken kayan aikin RNase-Free, babu buƙatar damuwa game da lalata RNA.
■ Rukunin Tsaftace DNA yana ɗaure musamman ga DNA, ta yadda kit ɗin zai iya cire gurɓataccen DNA ba tare da ƙara DNA ba.
∎ Yawan yawan amfanin RNA: Rukunin RNA-kawai da dabara na musamman na iya tsarkake RNA da kyau.
n Gudun sauri: mai sauƙin aiki kuma ana iya kammala shi cikin mintuna 30.
∎ Tsaro: ba a bužatar sinadarin reagents.
∎ Babban inganci: gutsuttssun RNA da aka tsarkake suna da tsafta, ba su da furotin da sauran ƙazanta, kuma suna iya saduwa da aikace-aikacen gwaji daban-daban.
Kit aikace-aikace
Ya dace da hakar da tsarkakewa na jimillar RNA daga samfuran nama mai sabo ko daskararre (musamman sabobin ganyayyakin ganyayyaki) tare da ƙaramin polysaccharide da abun ciki na polyphenol.
Gudun aiki
zane
Plant Total RNA Isolation Kit Plus ya sarrafa 50mg na sabobin ganyen polysaccharides da polyphenols, kuma an gwada 5% tsaftataccen RNA ta hanyar electrophoresis.
1: banana
2: ginkgo
3: auduga
4: Ruman
Adana da rayuwar shiryayye
Ana iya adana kit ɗin na tsawon watanni 12 a cikin ɗaki da zafin jiki (15-25 ℃) a bushe bushe, kuma 2-8 ℃ na tsawon lokaci (watanni 24).
Ana iya adana buffer PSL1 a 4 ℃ na wata 1 bayan ƙara 2-hydroxy-1-ethanethiol (na zaɓi).
Jagorar Binciken Matsala
Binciken na gaba na matsalolin da zaku iya fuskanta a cikiJimillar ShukaRNA extraction will help you with your experiments. In addition, for other experimental or technical problems in addition to operating instructions and problem analysis, we have dedicated technical support to help you. If you have any needs, please contact us at: 028-83360257 or E-mali : Tech@foregene.com.
Rukunin juyi yana toshe
Toshe ginshiƙin jujjuyawar zai haifar da rage yawan amfanin RNA ko ma kasa tsarkakewa don samun RNA, kuma ingancin RNA da aka samu zai yi ƙasa da ƙasa.
Nazarin Dalilan gama gari:
1. Samfurin ba a karye gaba daya ba.
Rarraba samfurin da bai cika ba zai iya toshe Rukunin Tsabtace DNA, wanda kuma zai iya shafar yawan amfanin RNA da inganci.Muna ba da shawarar cewa lokacin aiwatar da rarrabuwar samfur, da sauri niƙa cikin isassun ruwa na nitrogen don karya kyallen takarda kamar bangon tantanin halitta da membranes na samfuran gwargwadon yiwuwa.Don samfuran tsirrai na polyphenol polysaccharides, muna ba da shawarar ku yi amfani da Plant Total RNA Isolation Kit Plus.
2.Aspirate ginshiƙin Tsabtace DNA wanda ya keɓanta, yana neman yuwuwar tarkace tantanin halitta.
Pellet tarkacen tantanin halitta na iya toshe Rukunin RNA-kawai yayin hanyoyin tallan RNA (duba mataki na 5 na hanya, mataki na 6 na hanyar polysaccharide polyphenol).Muna ba da shawarar cewa dole ne a kula yayin da ake sha'awar wannan na'urar don gujewa tarkace tantanin halitta.
3. Adadin farko na samfurin ya yi yawa.
Yin amfani da samfurin da ya wuce kima zai haifar da rarrabuwar samfur mara cika ko rashin cikar lysis na sel ta Buffer PRL1 ko Buffer PSL1, yana haifar da ginshiƙin tsarkakewa mai toshe don ayyukan tsarkakewa.Kayan Tsire-tsire na keɓewar RNA yana da matsakaicin farko na 50 MG kowace tsarkakewa ɗaya na samfurin da aka sarrafa.Don samfuran tsirrai na polyphenol polysaccharides, muna ba da shawarar ku gwada Plant Total RNA Isolation Kit Plus.
4. Zazzabi na centrifuge yayi ƙasa da ƙasa.
Gabaɗayan keɓewar RNA da tsarkakewa ban da rushewar sinadarin nitrogen na ruwa, duk matakan ana yin su ne a cikin ɗaki (20-25 ° C).Wasu ƙananan zafin jiki na centrifuges suna da yanayin zafi ƙasa da 20 ° C, wanda zai iya haifar da toshewa a cikin Rukunin Tsabtace DNA da/ko Rukunin RNA-kawai.Idan wannan ya faru, saita zafin jiki na centrifuge zuwa 20-25 ° C da kuma dumi cakuda lysis da/ko ƙari na rabuwar ethanol zuwa 37 ° C.
Babu RNA da aka fitar ko RNA da ke ƙasa
Yawancin lokaci akwai dalilai da yawa waɗanda ke shafar ingancin farfadowa, kamar: samfurin abun ciki na RNA, hanyar aiki, ƙarar haske, da sauransu.
Binciken abubuwan gama gari kamar haka:
1.An yi wanka na kankara ko ƙananan zafin jiki (4 ° C) centrifugation a yayin aikin.
Shawarwari: Yi aiki a dakin da zafin jiki (15-25 ° C) a cikin dukan tsari, kada ku yi wanka na kankara da ƙananan zafin jiki.
2.An lalatar da RNA saboda rashin kyawun adana samfurin ko adana samfurin na dogon lokaci.
Shawarwari: Samfurori da aka tattara da sauri ya kamata a daskare su cikin ruwa nitrogen, sa'an nan kuma adana su a -80 ° C na dogon lokaci, guje wa daskarewa maimaituwa da narke samfurori;ko nan da nan jiƙa samfuran a cikin maganin RNAlater stabilizer (samfurori na dabba).
3.Rashin isasshen samfurin rarrabuwa da lysis yana haifar da toshe ginshiƙin tsarkakewa.
Shawara: Lokacin da ake niƙa nama, da fatan za a tabbatar cewa nama ya isa ƙasa, kuma da sauri canja shi zuwa PSL1 da aka riga aka shirya (tabbatar da cewa an ƙara daidai adadin β-ME, duba mataki na 1 na hanya).
4.An ƙara eluent ba daidai ba.
Shawara: Tabbatar da cewa ddH ba ta da RNase2O yana ɗigowa a tsakiyar membrane ginshiƙin tsarkakewa.
5.Ba a ƙara madaidaicin ƙarar cikakken ethanol zuwa Buffer PSL2 ko Buffer PRW2 ba.
Shawara: Da fatan za a bi umarnin, ƙara madaidaicin ƙarar cikakken ethanol zuwa Buffer PSL2 da Buffer PRW2 kuma a haɗe da kyau kafin a yi amfani da kit ɗin.
6.Yawan samfurin nama bai dace ba.
Shawara: Yi amfani da 50 MG na nama a cikin 500 μl na Buffer PSL1.Yin amfani da nama mai yawa zai rage adadin RNA da aka fitar sannan kuma za a rage tsaftar RNA da ke haifarwa.Muna ba da shawarar sosai cewa adadin samfurin farko bai kamata ya wuce 50 MG ba a kowace aikin cirewar RNA.
7. Ƙarfin ƙira mara dacewa ko rashin cikawa.
Shawarwari: Ƙaƙwalwar ƙarar ginshiƙin tsarkakewa shine 50-200 μl;idan tasirin elution bai gamsar ba, ana ba da shawarar tsawaita lokacin a cikin zafin jiki bayan ƙara ddH da aka rigaya ba RNase-Free.2O, kamar minti 5-10.
8.Shafin tsarkakewa yana da ragowar ethanol bayan wankewa tare da Buffer PRW2.
Shawarwari: Idan fanko bututu yana centrifuged na 1 min kuma har yanzu akwai sauran ethanol bayan wankewa a cikin Buffer PRW2, zaku iya ƙara lokacin faɗuwar bututun centrifugation zuwa 2 min, ko sanya ginshiƙin tsarkakewa a ɗakin zafin jiki na 5 min don cikakken cire ragowar ethanol.
9.An yi amfani da kit ɗin ba daidai ba.
Shawara: Don samfuran tsire-tsire na polysaccharides na polyphenolic, amfani da kayan gama gari kamar Plant Total RNA Isolation Kit maiyuwa ba zai iya samun ingantattun samfuran RNA ba.Muna ba ku shawarar amfani da Plant Total RNA IsolationKit Plus, wanda aka kera musamman don samfuran tsire-tsire na polyphenolic polysaccharide.Kit ɗin da aka haɓaka musamman don cire RNA daga samfuran tsirrai na polyphenol da polysaccharide.
Ƙimar OD260/OD280 tayi ƙasa
RNA haɓakawa tare da ddH2O kuma an yi amfani da shi don sakamakon karatun spectrophotometer a cikin ƙananan ƙimar OD260/OD280.Muna ba da shawarar yin amfani da 10 mM Tris-HCl, pH 7.5 (maimakon RNase-Free ddH2O don elute RNA) don samun ingantacciyar ƙimar OD260/OD280, duba "Tallafin RNA da Tsarkakewar Tsarkakewa" a shafi na 19.
RNA da aka tsarkake ta lalace
Ingancin RNA da aka tsarkake yana da alaƙa da abubuwa kamar adana samfur, gurɓatawar RNase, da magudi.
Binciken abubuwan gama gari:
1.Ba a adana samfuran nama a cikin lokaci bayan tattarawa.
Shawarwari: Idan ba a yi amfani da samfuran nama a cikin lokaci bayan tattarawa, da fatan za a adana su a cikin ruwa nitrogen a ƙananan zafin jiki nan da nan ko canza su zuwa -80 ° C don ajiya na dogon lokaci bayan daskarewa da sauri a cikin ruwa nitrogen, ko kuma nan da nan nutsar da samfuran a cikin maganin RNA stabilizer RNAlater (samfurori na dabba).Don hakar RNA, gwada amfani da samfuran nama da aka tattara sabo.
2.Maimaita daskarewa da narke samfuran nama.
Shawara: Lokacin adana samfuran nama, yana da kyau a yanke su kanana don adanawa, sannan a fitar da wani yanki daga cikinsu yayin amfani da su don guje wa lalatawar RNA da ke haifar da daskarewa akai-akai da narke samfurin.
An gabatar da 3.RNase a cikin dakin aiki ko ba a sawa safofin hannu, masks, da dai sauransu.
Shawara: An fi yin gwaje-gwajen hakar RNA a cikin ayyukan RNA daban-daban, kuma a tsaftace teburin dakin gwaje-gwaje kafin gwaji, kuma a sanya safar hannu da abin rufe fuska yayin gwajin don guje wa lalatawar RNA wanda ya haifar da gabatarwar RNase zuwa mafi girma.
4.The reagent yana gurbata ta RNase yayin amfani.
Shawara: Sauya tare da sabon jerin kayan aikin cirewar RNA gaba ɗaya don gwaje-gwaje masu alaƙa.
5.The centrifuge tubes da pipette tukwici da ake amfani da su don magudin RNA sun gurbata da RNase.
Shawarwari: Tabbatar cewa bututun centrifuge, tukwici na pipette, pipettes, da sauransu. da ake amfani da su wajen fitar da RNA duk ba su da RNase.
Jagoran Jagora:
