Animal Total RNA Isolation Kit Total RNA Extraction and Purification Kits for Animal Tissues & Cell
Kufotokozera Kwa Zida:
Kuchotsa mwachangu komanso moyenera RNA yapamwamba komanso yapamwamba kwambiri kuchokera kumagulu osiyanasiyana anyama.
Palibe chifukwa chodera nkhawa za kuwonongeka kwa RNA.Dongosolo lonse ndi RNase-Free
Chotsani bwino DNA pogwiritsa ntchito DNA-Cleaning Column
Chotsani DNA popanda kuwonjezera DNase
Zosavuta-zochita zonse zimatsirizidwa ndi kutentha kwa chipinda
Kuthamanga-ntchito ikhoza kutha m'mphindi 30
Otetezeka-palibe organic reagent yomwe imagwiritsidwa ntchito
Kuyera kwakukulu-OD260/280≈1.8-2.1
Mafotokozedwe a Kits
50 Preps, 200 Preps
Chida ichi chimagwiritsa ntchitospin column ndi formulayopangidwa ndi kampani yathu, yomwe imatha kuchotsa chiyero chapamwamba komanso chapamwamba kwambiri cha RNA kuchokera kumagulu osiyanasiyana a zinyama ndi ntchito yabwino kwambiri. Imapereka DNA-Cleaning Column yogwira mtima, yomwe ingathe kulekanitsa ndi kutsatsa DNA ya genomic kuchokera ku supernatant ndi lysate ya minofu, yosavuta komanso yopulumutsa nthawi;Mzere wa RNA wokha ukhoza kumangirira RNA bwino ndipo ukhoza kusinthidwa nthawi imodzi ndi chilinganizo chapadera Zitsanzo zambiri.
Dongosolo lonselo ndi RNase-Free, kotero kuti RNA yochotsedwayo isawonongeke;Buffer RW1, Buffer RW2 buffer makina ochapira, kotero kuti RNA yopezekayo ilibe mapuloteni, DNA, ion, ndi kuipitsidwa kwapawiri.
Zida zamagulu
| Animal Total RNA Isolation Kit | ||
| Zida zamagulu | RE-03011 | RE-03014 |
| 50 T | 200 T | |
| Buffer RL1* | 25ml ku | 100 ml |
| Mtengo wa RL2 | 15ml ku | 60 ml pa |
| Bafa RW1* | 25ml ku | 100 ml |
| Mtengo wa RW2 | 24ml ku | 96ml pa |
| RNase-Free ddH2O | 10 ml pa | 40 ml pa |
| Mzere wa RNA wokha | 50 | 200 |
| DNA-Kuyeretsa Mzere | 50 | 200 |
| Buku Lolangiza | 1 chidutswa | 1 chidutswa |
Zambiri zamalonda
| Mtundu | Mzere wozungulira | Chiyeretso gawo | Foregene column, reagent |
| Flux | 1-24 zitsanzo | Nthawi yokonzekera | ~30 min (24 zitsanzo) |
| Centrifuge | Desk centrifuge | Kupatukana kwa pyrolysis | Kupatukana kwa Centrifugal |
| Chitsanzo | Minofu ya nyama;selo | Zitsanzo kuchuluka | Minofu: 10-20 mg;Selo:(1-5)×106 |
| Elution voliyumu | 50-200 μL | Voliyumu yokweza kwambiri | 850 μl |
Mbali & ubwino
■ Palibe chifukwa chodera nkhawa za kuwonongeka kwa RNA;dongosolo lonse ndi RNase-Free
■ Chotsani bwino DNA pogwiritsa ntchito DNA-Cleaning Column
■ Chotsani DNA popanda kuwonjezera DNase
■ Zosavuta-zochita zonse zimatsirizidwa pa kutentha kwa chipinda
■ Kuthamanga kwachangu kumatha kutha mphindi 30
■ Safe-palibe organic reagent chofunika
■ Kuyera kwakukulu -OD260/280≈1.8-2.1
Kit application
Ndiwoyenera kuchotsedwa ndi kuyeretsedwa kwa RNA yonse kuchokera kumitundu yosiyanasiyana yanyama yatsopano kapena yozizira kapena maselo otukuka.
Zogulitsa katundu
■ Ntchito zotsika: kaphatikizidwe ka cDNA yoyamba, RT-PCR, kupanga maselo, Northern Blot, etc.
■ Zitsanzo: minofu ya nyama, maselo otukuka
■ Mlingo: Tissues 10-20mg, Maselo(2-5)×106
■ Kuchuluka kwa DNA kumangiriza mphamvu yoyeretsa: 80 μg
■ Voliyumu yowonjezera: 50-200 μl
Chithunzi
Animal Total RNA Isolation Kit ankachitira 20mg
Zitsanzo za mbewa zatsopano, tengani 5% oyeretsedwa okwana RNA 1% agar
Glycogel electrophoresis
1: Nkhumba 2: Impso
3: Chiwindi 4: Mtima
Kusungirako ndi moyo wa alumali
Zidazi zitha kusungidwa kwa miyezi 24 kutentha (15-25 ℃) kapena 2-8 ℃ kwa nthawi yayitali.Buffer RL1 ikhoza kusungidwa pa 4 ℃ kwa mwezi wa 1 mutawonjezera β- mercaptoethanol (posankha).
Nkhani zotchulidwa
1.NGA:18.808:Zheng, Q., Qin, F., Luo, R., et al.mRNA-Loaded Lipid-Monga Nanoparticles kwa Chiwindi Base Editing Kudzera Kukhathamiritsa kwa Central gulu Design.Adv.Ntchito.Mater.2021, 31, 2011068.doi:10.1002/adfm.202011068.
2.NGATI:18.187:He X, Hong W, Yang J, et al.Apoptosis yodziwikiratu yama cell ochiritsira tsinde yokonzekera imakhala ndi zotsatira zoteteza thupi kutulutsa phosphatidylserine.Signal Transduct Target Ther.2021 Jul 14; 6 (1): 270.doi: 10.1038/s41392-021-00688-z.
3.NGATI: 17.97:Dai Z, Liu H, Liao J, et al.Kusintha kwa N7-Methylguanosine tRNA kumawonjezera kumasulira kwa oncogenic mRNA komanso kumathandizira kupita patsogolo kwa intrahepatic cholangiocarcinoma.Mol cell.2021 Jul 29:S1097-2765(21)00555-4.doi: 10.1016/j.molcel.2021.07.003.
4.NGATI:9.225:Cao X, Shu Y, Chen Y, et al.Kusintha kwa Mettl14-Mediated m6A Kumathandizira Kusinthika kwa Chiwindi Posunga Endoplasmic Reticulum Homeostasis.Cell Mol Gastroenterol Hepatol.2021;12(2):633-651.doi: 10.1016/j.jcmgh.2021.04.001.
RNA isoaltion zida za zina zitsanzo magwerozilipo:
Selo, zomera, tizilombo, magazi, etc.
RNA sinachotsedwe kapena zokolola za RNA ndizochepa
Nthawi zambiri pamakhala zinthu zosiyanasiyana zomwe zimakhudza kuchira bwino, monga: minofu ya RNA yokhutira, njira yogwirira ntchito, voliyumu ya elution, ndi zina zambiri.
1. Kusambira kwa ayezi kapena cryogenic (4 ° C) centrifugation kunachitika panthawi ya ntchito.
Malangizo: Gwiritsani ntchito kutentha (15-25 ° C) panthawi yonseyi, osasamba madzi oundana ndi centrifuge pa kutentha kochepa.
2. Kusungidwa kwachitsanzo kosayenera kapena nthawi yochuluka yosungira zitsanzo.
Malangizo: Sungani zitsanzo pa -80 ° C kapena kuzizira mu nayitrogeni wamadzimadzi ndipo pewani kugwiritsa ntchito kuzizira kobwerezabwereza;yesetsani kugwiritsa ntchito minofu yatsopano kapena maselo otukuka kuti muchotse RNA.
3. Kusakwanira kwachitsanzo lysis.
Malangizo: Mukamagwirizanitsa minofu, onetsetsani kuti minofuyo ili ndi homogenized mokwanira komanso kuti maselo a minofu agawanika mokwanira kuti afotokoze kutulutsidwa kwa RNA.
4. Chosowacho sichiwonjezedwa bwino.
Malangizo: Tsimikizirani kuti RNase-Free ddH2O amawonjezedwa pang'onopang'ono pakati pa nembanemba yoyeretsa.
5. Voliyumu yolondola ya ethanol yokwanira sinawonjezedwe ku Buffer RL2 kapena Buffer RW2.
Malangizo: Tsatirani malangizo, onjezani voliyumu yolondola ya ethanol ku Buffer RL2 ndi Buffer RW2 ndikusakaniza bwino musanagwiritse ntchito zida.
6. Mlingo wa zitsanzo za minofu siwoyenera.
Malangizo: Gwiritsani ntchito 10-20 mg ya minofu kapena (1-5) × 106ma cell pa 500 μl buffer RL1, chifukwa kugwiritsa ntchito minofu mopitirira muyeso kungayambitse kuchepa kwa RNA.
7. Voliyumu yolakwika kapena kusakwanira.
Malangizo: Voliyumu ya elution ya gawo loyeretsa ndi 50-200 μl;ngati mphamvu ya elution siili yokhutiritsa, tikulimbikitsidwa kuwonjezera nthawi yoyika kutentha kwa chipinda mutawonjezera RNase-Free ddH yotenthetsera.2O, mwachitsanzo kwa 5-10 min.
8.Mzere woyeretsedwa uli ndi zotsalira za ethanol pambuyo pa kusamba kwa Buffer RW2.
Malangizo: Ngati pali zotsalira za ethanol pambuyo pa kusamba kwa Buffer RW2, chubu chopanda kanthu centrifugation kwa 1min, nthawi yopanda kanthu chubu centrifugation ntchito akhoza ziwonjezeke kwa 2min, kapena ndime kuyeretsa akhoza kuikidwa pa firiji kwa mphindi 5 mokwanira kuchotsa Mowa yotsalira.
RNA yoyeretsedwa ndiyonyozeka
Ubwino wa RNA yoyeretsedwa ikugwirizana ndi zinthu monga kusungidwa kwa chitsanzo, kuipitsidwa kwa RNase, ndi kusintha, ndi zina zotero.
1. Zitsanzo za minofu sizimasungidwa nthawi.
Malangizo: Ngati zitsanzo za minofu kapena maselo sagwiritsidwa ntchito munthawi yake mutatolera, nthawi yomweyo sungani pa -80 °C kapena nayitrogeni wamadzimadzi.Kuti mutulutse RNA, gwiritsani ntchito minofu yongotengedwa kumene kapena maselo a cell ngati kuli kotheka.
2. Kuzizira kobwerezabwereza kwa zitsanzo za minofu.
Malangizo: Posunga zitsanzo za minofu, ndi bwino kuzidula kuti zisungidwe, ndikuchotsa chimodzi mwa zidutswazo pozigwiritsira ntchito kuti musamaundane mobwerezabwereza ndi kuwonongeka kwa RNA.
3. RNase imayambitsidwa kapena kusavala magolovesi otayika, masks, ndi zina zotero panthawi ya opaleshoni.
Malangizo: Kuyesa kwa RNA m'zigawo kumachitika bwino m'zipinda zosinthira za RNA ndipo tebulo limachotsedwa kuyesako kusanachitike.
Valani magolovesi ndi masks otayika panthawi yoyesera kuti muchepetse kuwonongeka kwa RNA komwe kumachitika chifukwa cha kukhazikitsidwa kwa RNase.
4. Ma reagents amadetsedwa ndi RNase pakugwiritsa ntchito.
Malangizo: M'malo ndi Animal Total RNA Isolation Kit yatsopano yoyeserera yofananira.
5. Machubu a centrifuge, nsonga, ndi zina zotere zomwe zimagwiritsidwa ntchito pakusintha kwa RNA zimaipitsidwa ndi RNase.
Malangizo: Tsimikizirani kuti machubu a centrifuge, nsonga, mapaipi, ndi zina zomwe zimagwiritsidwa ntchito pochotsa RNA zonse ndi Zaulere za RNase.
Oyeretsedwa analandira RNA amakhudza zoyesera kumunsi
RNA yoyeretsedwa ndi ndime yoyeretsera, ngati ayoni amchere, zomanga thupi ndi zazikulu kwambiri zidzakhudza kuyesera kumtunda, monga: reverse transcription,Northern Blot et al.
1. The elutioned RNA ili ndi zotsalira za ayoni zamchere.
Malangizo: Tsimikizirani kuti voliyumu yolondola ya ethanol yawonjezedwa ku Buffer RW2 ndikuchita ndime zoyeretsera 2 pa liwiro la centrifugal lomwe lasonyezedwa kuti ligwire ntchito;ngati pali zotsalira za mchere wa ion, siyani mzere woyeretsera ku Buffer RW2 kwa mphindi 5 pa kutentha kwapakati ndikuchita centrifugation kuti muwonjezere kuchotsa kuipitsidwa kwa mchere.
2. Ethanol zotsalira mu elutioned RNA.
Malangizo: Tsimikizirani kuti mutatha kutsuka kwa bafa RW2, gwiritsani ntchito chubu cha centrifugation chopanda kanthu pa liwiro la centrifugation lomwe likuwonetsedwa kuti ligwire ntchito, onjezerani nthawi ya ntchito yopanda kanthu ya chubu centrifugation mpaka 2 min ngati pali zotsalira za ethanol, kapena musiye kutentha kwa mphindi 5 pambuyo pa chubu chopanda kanthu centrifugation kuti muwonjezere kuchotsedwa kwa ethanol.
