Jumlar Tsirrai RNA Kit ɗin Keɓewa Tare da Jimlar RNA Purificaiton Kit don Mai Wadatar Shuka a cikin Polysaccharides da Polyphenols
Bayanin Kit:
Cat.No.RE-05021/05022/05024
Don tsarkakewa na jimlar RNA daga samfuran tsire-tsire na gabaɗaya waɗanda ke ɗauke da manyan polysaccharide da abubuwan polyphenol.
Cire RNA mai inganci da sauri daga samfuran shuka tare da babban abun ciki na polysaccharides da polyphenols.
RNase-Free Amfani da Rukunin Tsabtace DNA
Sauƙaƙe-dukkan ayyukan ana kammala su a cikin zafin jiki
Ana iya kammala aiki mai sauri a cikin mintuna 30
Amintacciya — ba a yi amfani da reagent Organic ba 
Ƙayyadaddun bayanai
Shirye-shirye 50, Shirye-shiryen 200
Kit ɗin yana amfani da ginshiƙin juzu'i da dabara wanda Foregene ya haɓaka, wanda zai iya fitar da inganci mai inganci da ingantaccen jimlar RNA daga kyallen shuka iri-iri tare da babban polysaccharides ko abun ciki na polyphenols.Yana ba da ginshiƙin Tsabtace DNA wanda zai iya cire DNA na genomic cikin sauƙi 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 |
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.
Musamman dacewa don tsarkakewar RNA daga samfuran shuka na polysaccharides da polyphenols.
■ 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.
Siffofin samfur
∎ Aikace-aikace na ƙasa: haɗin cDNA na farko, RT-PCR, cloning na kwayoyin halitta, Northern Blot, da sauransu.
Misali: Sabbin kyallen tsire-tsire ko daskararre na polysaccharides da polyphenols
■ Adadin: 50mg nama na shuka
n Matsakaicin ƙarfin ɗaurin RNA na ginshiƙin tsarkakewa: 80 μg
■ Ƙarfin haske: 50-200 μl
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 babban 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 Shelf
Ana iya adana wannan kit ɗin na tsawon watanni 24 a ƙarƙashin busassun yanayi a cikin zafin jiki (15-25 ℃);Idan ana buƙatar adana na dogon lokaci, ana iya adana shi a cikin 2-8 ℃.
Buffer PSL1 za a iya sanya shi a 4 ℃ na wata 1 bayan ƙara β-mercaptoethanol (an bada shawarar ƙara shi a lokaci guda na gwaji).
Rukunin ya toshe
Bayan an toshe ginshiƙi, yawan amfanin RNA yana raguwa ko ma ba zai yiwu ba don tsarkake RNA, kuma adadin RNA da aka samu yayi ƙasa.
Bincike na gama gari:
1. Samfurin karya ba cikakke ba ne.
Karyewar samfurin ba ya haifar da toshewar ginshiƙi na DNA gaba ɗaya, yayin da yake shafar yawan amfanin RNA da inganci.Muna ba da shawarar aikin niƙa da sauri a cikin isasshen ruwa nitrogen lokacin da kuka karya samfuran, Yi ƙoƙarin murkushe bangon samfurin tantanin halitta, membrane cell da sauran nama.Don samfuran tsire-tsire na polyol polysaccharides, muna ba da shawarar ku yi amfani da Plant Total RNA ISOLATION KIT PLUS.
2. Lokacin da tsotsan samfurin da aka keɓe tare da ginshiƙin Tsabtace DNA, yuwuwar guguwar tantanin halitta na iya shakar.
Rukunin ɓangarorin tantanin halitta da aka ɗauka zai haifar da Rukunin RNA-ONLY wanda za a toshe lokacin da aka yi aikin tallan RNA (duba mataki na 6).Muna ba ku shawarar a hankali lokacin da ake tsotsa wannan abin sha don guje wa tsotsar tarkace ta cell.
3. Samfurin adadin farko yayi yawa.
Yin amfani da samfurin da ya wuce kima zai haifar da raguwar samfurin da bai cika ba ko kuma rashin cikar lysis ta Buffer PSL1, yana haifar da toshe ginshiƙin tsarkakewa yayin tsarkakewa.Jumlar Shuka Kayan Keɓewar RNA Kowane samfurin aikin da aka tsarkake guda ɗaya shine MG 50.Don samfuran tsirrai na polyol polysaccharides, muna ba da shawarar ku gwada Plant Total RNA ISOLATION KIT PLUS.
4. Zazzabi na centrifuge yayi ƙasa da ƙasa.
Ana aiwatar da duk keɓewar RNA da tsarin tsarkakewa a cikin zafin jiki (20-25°C), sai dai cewa samfurin nama ya karye ta hanyar ruwa nitrogen. Zazzabi na wasu centrifuges cryogenic yana ƙasa da 20℃, wanda zai iya haifar da toshe Rukunin Tsabtace DNA da/ko Rukunin RNA-Only.Idan wannan ya faru, saita zafin jiki na centrifuge zuwa 20-25℃, kumaTabbatar cewa cakudawar lysis da/ko an riga an riga an riga an riga an yi zafi da sinadarin 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 da zafin jiki (15-25°C) a cikin dukan tsari, kada ku yi wanka na kankara da ƙananan zafin jiki centrifugation.
2.An lalata RNA saboda rashin daidaituwa na samfurin ko adana dogon lokaci na samfurin.
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 cewa ddH2O-Free RNase ya ɗigo zuwa tsakiyar ginshiƙi mai 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.Rashin da bai dace ba ko kuma 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 ddH2O da aka rigaya da RNase-Free, kamar 5-10min.
8.Shafin tsarkakewa yana da ragowar ethanol bayan wankewa tare da BufferPRW2.
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 elution tare da ddH2O kuma ana 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 haɓaka RNA) don samun ingantacciyar ƙimar OD260/OD280, duba "Takaddar Tattaunawa da Tsarkakewar RNA" 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:
Shuka Jimlar RNA Keɓewar Kit Plus Jagoran Umarni
