Cell Total RNA Isolation Kit Total RNA Isolation Purification Kits kuchokera mu selo
Kufotokozera Kwa Zida:
RNA yoyeretsedwa kwambiri komanso yapamwamba kwambiri imatha kupezeka m'maselo osiyanasiyana azikhalidwe m'mphindi 11.
RNase-Free
Imagwiritsidwa ntchito kutentha (15-25 ℃)
Ndi DNA-Cleaning Column
Mtengo wapatali wa magawo RNA
Fast: Malizitsani kutulutsa mu mphindi 11
Chitetezo: Palibe Organic Chemical
Kufotokozera
Chidachi chimagwiritsa ntchito spin column ndi formula yopangidwa ndi Foregene, yomwe imatha kutulutsa RNA yapamwamba komanso yapamwamba kwambiri kuchokera ku ma cell opangidwa mu mbale 96, 24, 12, ndi 6-zitsime.
Chidachi chimapereka DNA-Cleaning Column yogwira mtima, yomwe imatha kulekanitsa ma supernatant ndi cell lysate, kumanga ndi kuchotsa DNA ya genomic.Ntchitoyi ndi yosavuta komanso yopulumutsa nthawi.
Tiye RNA-Column yokha imatha kumangirira RNA ndi njira yapadera.Zitsanzo zambiri zimatha kukonzedwa nthawi imodzi.
Zida zamagulu
| Kupanga kwa zida | RE-03111 | RE-03114 |
| 50 T | 200 T | |
| Buffer cRL1* | 25 ml pa | 100 ml |
| Mtengo wa cRL2 | 15 ml pa | 60 ml pa |
| Bafa RW1* | 25 ml pa | 100 ml |
| Mtengo wa RW2 | 24 ml pa | 96 ml pa |
| RNase-Free ddH2O | 10 ml pa | 40 ml pa |
| RNA-Only Column | 50 | 200 |
| DNA-Kuyeretsa Mzere | 50 | 200 |
| Malangizo | 1 | 1 |
*Chonde valani magolovesi ndikuchitapo kanthu zodzitchinjiriza mukamagwira ntchito popeza Buffer cRL1 ndi Buffer RW1 zili ndi mchere wachaotropic wokwiyitsa.
Mbali & ubwino
■ Njira yonseyi imayendetsedwa ndi kutentha kwa chipinda (15-25 ℃), popanda kusamba kwa ayezi ndi kutentha kochepa kwa centrifugation.
■ Zida zonse ndi RNase-Free, palibe chifukwa chodera nkhawa za kuwonongeka kwa RNA.
■ DNA-Cleaning Column imamangiriza DNA, kotero kuti zida zitha kuchotsa kuipitsidwa kwa DNA popanda kuwonjezera DNase.
■ Zokolola zambiri za RNA: Mzere wa RNA-okha ndi fomula yapadera imatha kuyeretsa RNA bwino.
■ Kuthamanga kwachangu: kosavuta kugwiritsa ntchito ndipo kumatha kumalizidwa mu mphindi 11.
■ Chitetezo: Palibe organic reagent yomwe imafunikira.
■ Ubwino wapamwamba: RNA yoyeretsedwa ndi yoyera kwambiri, yopanda mapuloteni ndi zonyansa zina, ndipo imatha kukumana ndi mayesero osiyanasiyana.
Kit application
Ndiwoyenera kuchotsa ndi kuyeretsa RNA yonse kuchokera ku maselo otukuka mu mbale 96, 24, 12, ndi 6-chitsime.
Kuyenda kwa ntchito
Chithunzi
The agarose gel osakaniza batire chithunzi cha Cell Total RNA Isolation Kit ankachitira pamwamba manambala osiyanasiyana a maselo, 20μl voliyumu elution, kutenga 2μl oyeretsedwa okwana RNA 1%.
Kusungirako ndi moyo wa alumali
Zida zitha kusungidwa kwa miyezi 12 kutentha kwa chipinda (15-25 ℃) kapena 2-8 ℃ kwa nthawi yayitali (miyezi 24).
Buffer cRL1 ikhoza kusungidwa pa 4 ℃ kwa mwezi umodzi mutawonjezera 2-hydroxy-1-ethanethiol (posankha).
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 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 5 m.
