Mhuka Yese RNA Isolation Kit Yese RNA Kubvisa uye Kuchenesa Kits yeMhuka Tissues & Sero
Kit Description:
Nekukurumidza uye zvinobudirira bvisa yakakwira-kuchena uye yemhando yepamusoro RNA kubva kune akasiyana mhuka tishu.
Hapana chikonzero chekunetseka nezveRNA kuderedzwa.Iyo yese system ndeye RNase-Yemahara
Kunyatso bvisa DNA uchishandisa DNA-Cleaning Column
Bvisa DNA pasina kuwedzera DNase
Zviri nyore-mabasa ese anopedzwa patembiricha yekamuri
Kukurumidza-kushanda kunogona kupedzwa mumaminitsi makumi matatu
Yakachengeteka-hapana organic reagent inoshandiswa
Kuchena kwepamusoro-OD260/280≈1.8-2.1
Kits Tsanangudzo
50 Preps, 200 Preps
Iyi kit inoshandisa iyospin column uye formulayakagadzirwa nekambani yedu, iyo inogona kubvisa yakakwirira-kuchena uye yepamusoro-soro yakazara RNA kubva kumarudzi akasiyana-siyana emhuka nepamusoro-soro.Inopa DNA-Cleaning Column inoshanda, iyo inogona kuparadzanisa nyore nyore uye adsorb genomic DNA kubva kune supernatant uye tissue lysate, iri nyore uye inochengetedza nguva;RNA-chete Column inokwanisa kusunga RNA zvakanaka uye inogona kugadziriswa panguva imwe chete neyakasarudzika fomula Mazhinji emasamples.
Iyo yese sisitimu ndeye RNase-Yemahara, kuitira kuti iyo yakabviswa RNA irege kuderedzwa;Buffer RW1, Buffer RW2 buffer washing system, kuitira kuti iyo RNA inowanikwa haina mapuroteni, DNA, ion, uye organic compound kusvibiswa.
Kit zvikamu
| Mhuka Yese RNA Isolation Kit | ||
| Kit zvikamu | RE-03011 | RE-03014 |
| 50 T | 200 T | |
| Buffer RL1* | 25ml | 100ml |
| Buffer RL2 | 15ml | 60ml |
| Buffer RW1* | 25ml | 100ml |
| Buffer RW2 | 24ml | 96ml |
| RNase-Yemahara ddH2O | 10ml | 40ml |
| RNA-chete Column | 50 | 200 |
| DNA-Kuchenesa Column | 50 | 200 |
| Bhuku reMirayiridzo | 1 chidimbu | 1 chidimbu |
Product information
| Format | Spin column | Kucheneswa chikamu | Foregene column, reagent |
| Flux | 1-24 samples | Nguva yekugadzirira | ~30 min (24 samples) |
| Centrifuge | Desk centrifuge | Pyrolysis kupatsanurwa | Centrifugal separation |
| Muenzaniso | Animal tissue;sero | Samples mari | Muviri: 10-20 mg;Sero:(1-5)×106 |
| Elution volume | 50-200 μL | Maximum loading volume | 850 μL |
Zvimiro&zvakanakira
■ Hapana chikonzero chekunetseka pamusoro pekuparara kweRNA;iyo yese system ndeye RNase-Yemahara
■ Bvisa DNA-uchishandisa DNA-Cleaning Column
■ Bvisa DNA pasina kuwedzera DNase
■ Kushanda kuri nyore-zvose zvinopedzwa patembiricha yepakamuri
■ Kukurumidza-kushanda kunogona kupedzwa mumaminitsi makumi matatu
■ Yakachengeteka-hapana organic reagent inodiwa
■ Kuchena kwepamusoro -OD260/280≈1.8-2.1
Kit application
Inokodzera kudhirowa uye kucheneswa kweiyo RNA yakazara kubva kune akasiyana siyana matsva kana akaomeswa mhuka mhuka kana maseru ane tsika.
Product parameters
■ Zvishandiso zvepasi: yekutanga-strand cDNA synthesis, RT-PCR, molecular cloning, Northern Blot, nezvimwe.
∎ Mienzaniso: matishu emhuka, masero ane tsika
■ Dosage: Matishu 10-20mg, Masero(2-5)×106
■ Maximum DNA inosunga simba rekuchenesa column: 80 μg
■ Elution volume: 50-200 μl
Mufananidzo
Mhuka Yese RNA Isolation Kit yakarapwa 20mg
Fresh mbeva samples, tora 5% yakacheneswa yakazara RNA 1% agar
Glycogel electrophoresis
1: Spleen 2: itsvo
3: Chiropa 4: Mwoyo
Kuchengetedza uye hupenyu hwesherufu
Iyo kit inogona kuchengetwa kwemwedzi makumi maviri nemana pamhepo tembiricha (15-25 ℃) kana 2-8 ℃ kwenguva yakareba.Buffer RL1 inogona kuchengetwa pa4 ℃ kwemwedzi 1 mushure mekuwedzera β- mercaptoethanol (kusarudza).
Nyaya dzakataurwa
1.IF:18.808:Zheng, Q., Qin, F., Luo, R., et al.mRNA-Yakatakura Lipid-Senge Nanoparticles yeLiver Base Editing kuburikidza neKugadziridza kweCentral Composite Design.Adv.Funct.Mater.2021, 31, 2011068.doi:10.1002/adfm.202011068.
2.IF:18.187:Iye X, Hong W, Yang J, nevamwe.Spontaneous apoptosis yemasero mune yekurapa stem cell kugadzirira inoshandisa immunomodulatory mhedzisiro kuburikidza nekuburitswa kwephosphatidylserine.Signal Transduct Target Ther.2021 Jul 14;6(1):270.doi: 10.1038/s41392-021-00688-z.
3.IF:17.97:Dai Z, Liu H, Liao J, nevamwe.N7-Methylguanosine tRNA kugadziriswa kunowedzera oncogenic mRNA shanduro uye inosimudzira intrahepatic cholangiocarcinoma kufambira mberi.Mol Cell.2021 Jul 29:S1097-2765(21)00555-4.doi: 10.1016/j.molcel.2021.07.003.
4.KANA:9.225:Cao X, Shu Y, Chen Y, et al.Mettl14-Mediated m6A Kugadziriswa Kunofambisa Chiropa Kuberekwazve neKuchengetedza Endoplasmic Reticulum Homeostasis.Sero Mol Gastroenterol Hepatol.2021;12(2):633-651.doi: 10.1016/j.jcmgh.2021.04.001.
RNA isoaltion kits ye mamwe sampuli masosizviripo:
Sero, chirimwa, utachiona, ropa, zvichingodaro.
RNA haina kubviswa kana RNA goho rakaderera
Panowanzove nemhando dzakasiyana dzezvinhu zvinokanganisa kudzoreredza kushanda zvakanaka, senge: tishu sampuli RNA yemukati, nzira yekushanda, elution vhoriyamu, nezvimwe.
1. Ice bath kana cryogenic (4 ° C) centrifugation yakaitwa panguva yekushanda.
Kurudziro: Shanda pamhepo tembiricha (15-25 ° C) mukati mese muitiro, usaite ice yekugezesa uye centrifuge pane yakaderera tembiricha.
2. Kuchengetedzwa kwemuenzaniso kusina kukodzera kana nguva yakawandisa yekuchengetedza sampuli.
Kurudziro: Chengetedza samples pa -80 °C kana kuomesa munitrogen yemvura uye dzivirira kudzokororwa kushandiswa kwechando-nyoro;edza kushandisa matishu matsva kana maseru akagadzirwa eRNA kutorwa.
3. Kusakwana sampuli lysis.
Kurudziro: Paunenge uchigadzira matishu, ita shuwa kuti matishu akanyatsoenderana uye kuti masero ematishu akapatsanurwa zvakakwana kutsanangura kuburitswa kweRNA.
4. Iyo enzi haina kuwedzerwa nemazvo.
Kurudziro: Simbisa kuti RNase-Yemahara ddH2O inowedzerwa kudonha kusvika pakati peyekucheneswa column membrane.
5. Vhoriyamu chaiyo yeabsolute ethanol haina kuwedzerwa kuBuffer RL2 kana Buffer RW2.
Kurudziro: Tevedzera mirairo, wedzera iyo chaiyo vhoriyamu yeabsolute ethanol kuBuffer RL2 uye Buffer RW2 uye sanganisa zvakanaka usati washandisa kiti.
6. Tissue sample dosage haina kukodzera.
Kurudziro: Shandisa 10-20 mg yetishu kana (1-5) × 106masero pa 500 μl buffer RL1, sezvo kushandiswa kwematishu zvakanyanya kunogona kukonzera kuderedzwa kweRNA.
7. Vhoriyamu yakashata isina kunaka kana kuti kuburitsa kusina kukwana.
Kurudziro: Vhoriyamu yekujekesa yekoramu yekuchenesa ndeye 50-200 μl;kana iyo elution mhedzisiro isingagutsi, zvinokurudzirwa kuwedzera tembiricha yekuisa yekamuri nguva mushure mekuwedzera preheated RNase-Mahara ddH.2O, eg kwe5-10 min.
8.Koramu yekuchenesa ine ethanol yakasara mushure meBuffer RW2 wash.
Kurudziro: Kana paine ethanol yakasara mushure mekugezwa kweBuffer RW2, isina chinhu chubhu centrifugation ye1min, nguva yekushayikwa kwechubhu centrifugation operation inogona kuwedzerwa kusvika 2min, kana mbiru yekuchenesa inogona kuiswa pakushambidzika kwemaminetsi mashanu kuti ibvise zvakakwana ethanol yakasara.
Yakanatswa RNA yakaderedzwa
Hunhu hweRNA yakacheneswa hune hukama nezvinhu zvakaita sekuchengetedza sampuli, kusvibiswa kweRNase, uye kunyengera, nezvimwe.
1. Tissue samples haisi kuchengetwa munguva.
Kurudziro: Kana masamples ematishu kana maseru akasashandiswa nenguva mushure mekuunganidzwa, pakarepo cryopreserve pa -80 ° C kana liquid nitrogen.Kuti ubvise RNA, shandisa ichangobva kutorwa tishu kana sero sampuli pese pazvinogoneka.
2. Kudzokororwa kwechando-kunyunguduka kwemasampuli ematishu.
Kurudziro: Paunenge uchichengeta matishu samples, zviri nani kuacheka kuita zvidimbu zvidiki kuti zvichengetedzwe, uye kubvisa chimwe chezvidimbu paunenge uchizvishandisa kudzivirira kudzokororwa nechando-kunyunguduka kwesample uye kushatiswa kweRNA.
3. RNase inounzwa kana kusapfeka magirovhosi anoraswa, masiki, nezvimwewo panguva yekushanda.
Kurudziro: RNA yekuwedzera yekuedza inoitwa zvakanyanya mumakamuri akasiyana eRNA manipulation uye tafura inocheneswa isati yaedzwa.
Pfeka magurovhosi anoraswa uye masks panguva yekuyedza kuderedza kuderedzwa kweRNA kunokonzerwa nekuunzwa kweRNase.
4. Zvigadzirwa zvakasvibiswa neRNase panguva yekushandiswa.
Kurudziro: Tsiva neMhuka Yakazara RNA Isolation Kit yezviyedzo zvinoenderana.
5. Machubhu ecentrifuge, mazano, nezvimwewo zvinoshandiswa muRNA manipulation zvakasvibiswa neRNase.
Kurudziro: Simbisa kuti machubhu ecentrifuge, matipi, mapipette, nezvimwewo anoshandiswa muRNA kudhirowa ese RNase-Mahara.
Yakacheneswa yakawanikwa RNA inokanganisa kuyedza kwepasi
RNA yakacheneswa nekoramu yekuchenesa, kana maion emunyu, mapuroteni akawandisa anozokanganisa kuyedza kwakadzika, senge: reverse transcription,Northern Blot et al.
1. Iyo elutioned RNA ine salt ion residues.
Kurudziro: Simbisa kuti vhoriyamu chaiyo ye ethanol yawedzerwa kuBuffer RW2 uye ita 2 mbiru yekunatsa washes pa centrifugal speed inoratidzwa kuti ishande;kana paine ion inosara yemunyu, siya mbiru yekuchenesa kuBuffer RW2 kwemaminetsi mashanu patembiricha yekamuri uye ita centrifugation kuti uwedzere kubviswa kwemunyu.
2. Ethanol yakasara mune elutioned RNA.
Kurudziro: Simbisa kuti mushure mekutsvaira RW2 kushambidza, ita isina chinhu tube centrifugation operation pa centrifugation speed inoratidzwa kuti ishande, wedzera nguva yekushanda isina chinhu chubhu centrifugation kusvika 2 min kana ichine ethanol yakasara, kana kuisiya pane imwe tembiricha ye 5 min mushure mokunge isina chinhu chubhu centrifugation kuti iwedzere kubviswa kwe ethanol.
