Chomera Chokwanira cha RNA Isolation Kit Total RNA Purificaiton Kit ya Zomera Zochepa mu Polysaccharides ndi Polyphenols
Zofotokozera
50 Preps, 200 Preps
Chidacho chimagwiritsa ntchito spin column ndi formula yopangidwa ndi Foregene, yomwe imatha kuchotsa bwino RNA yoyera komanso yapamwamba kwambiri kuchokera kumitundu yosiyanasiyana yazomera yokhala ndi ma polysaccharides otsika ndi ma polyphenols.Pazitsanzo za zomera zokhala ndi ma polysaccharides apamwamba kapena ma polyphenols, tikulimbikitsidwa kugwiritsa ntchito Plant Total RNA Isolation Plus Kit kuti mupeze zotsatira zabwinoko zochotsa RNA.Chidachi chimapereka DNA-Cleaning column yomwe imatha kuchotsa mosavuta DNA ya genomic kuchokera ku supernatant ndi minofu lysate.Mzere wa RNA wokha ukhoza kumangirira RNA.Chidachi chimatha kukonza zitsanzo zambiri nthawi imodzi.
Dongosolo lonse lilibe RNase, kotero RNA yoyeretsedwa sidzanyozedwa.Buffer PRW1 ndi Buffer PRW2 akhoza kuonetsetsa kuti RNA yopezedwa siipitsidwa ndi mapuloteni, DNA, ayoni, ndi organic compounds.
Zida zamagulu
| Buffer PSL1, Buffer PS, Buffer PSL2 |
| Buffer PRW1, Buffer PRW2 |
| RNase-Free ddH2O, Mzere Woyeretsa DNA |
| RNA-Only Column |
| Malangizo |
Mbali & ubwino
■ Opaleshoni firiji (15-25 ℃) lonse ndondomeko, popanda ayezi kusamba ndi otsika kutentha centrifugation.
■ Zida zonse za RNase-Free, palibe chifukwa chodera nkhawa za kuwonongeka kwa RNA.
■ DNA-Cleaning Column imamangiriza ku DNA, 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 kugwira ntchito ndipo kumatha kutha mkati mwa mphindi 30.
■ Chitetezo: palibe organic reagent yomwe imafunikira.
■ Ubwino wapamwamba: Zidutswa za RNA zoyeretsedwa ndi zoyera kwambiri, zopanda zomanga thupi ndi zonyansa zina, ndipo zimatha kukumana ndi zoyeserera zosiyanasiyana zakutsika.
Kit application
Ndiwoyenera kuchotsa ndi kuyeretsa RNA yonse kuchokera ku zitsanzo zamafuta a zomera zatsopano kapena zozizira (makamaka masamba atsopano a masamba) okhala ndi polysaccharide yochepa ndi polyphenol.
Kuyenda kwa ntchito
Chithunzi
Plant Total RNA Isolation Kit Plus inakonzedwa 50mg masamba atsopano a polysaccharides ndi polyphenols, ndipo 5% yoyeretsedwa RNA inayesedwa ndi electrophoresis.
1: nthochi
2: Chiko
3: thonje
4: Khangaza
Kusungirako ndi moyo wa alumali
Zida zimatha kusungidwa kwa miyezi 12 kutentha kwa firiji (15-25 ℃) pamalo owuma, ndi 2-8 ℃ kwa nthawi yayitali (miyezi 24).
Buffer PSL1 ikhoza kusungidwa pa 4 ℃ kwa mwezi umodzi mutawonjezera 2-hydroxy-1-ethanethiol(ngati mukufuna).
Vuto Kusanthula Buku
Kuwunika kotsatiraku kwamavuto omwe mungakumane nawoZomera ZonseRNA 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.
Mzere wozungulira watsekedwa
Kutsekeka kwa mzere wozungulira kumapangitsa kuti zokolola za RNA zichepe kapena kulephera kuyeretsedwa kuti mupeze RNA, ndipo mtundu wa RNA womwe wapezedwa udzakhala wotsika.
Kusanthula Zomwe Zimayambitsa:
1. Chitsanzo sichinaswe kwathunthu.
Kugawikana kosakwanira kwachitsanzo kumatha kuletsa DNA-Cleaning Column, yomwe ingakhudzenso zokolola za RNA ndi mtundu wake.Tikukulimbikitsani kuti mukamagawanitsa zitsanzo, perani mwachangu kuchuluka kwa nayitrogeni wamadzimadzi kuti muphwanye minyewa monga makoma a cell ndi nembanemba zama cell momwe mungathere.Pazitsanzo za mbewu za polyphenol polysaccharides, timalimbikitsa kuti mugwiritse ntchito Plant Total RNA Isolation Kit Plus.
2.Aspirate the DNA-Cleaning Column isolated supernatant, aspirate the zotheka cell zinyalala pellet.
Aspirated cell debris pellet imatha kutseka RNA-Oly Column panthawi ya RNA adsorption process (onani sitepe 5 ya ndondomeko, sitepe 6 ya polysaccharide polyphenol process).Tikukulimbikitsani kuti musamalire pamene mukulakalaka chodabwitsa ichi kuti mupewe zinyalala zama cell.
3. Chiwerengero choyambirira cha zitsanzo ndi chachikulu kwambiri.
Kugwiritsa ntchito kwambiri zitsanzo kumabweretsa kugawikana kosakwanira kwa ma cell kapena kusakwanira kwathunthu kwa maselo ndi Buffer PRL1 kapena Buffer PSL1, zomwe zimapangitsa kuti pakhale gawo lotsekeka loyeretsera ntchito zoyeretsa.Plant Total RNA Isolation Kit ili ndi kuchuluka koyambirira kwa 50 mg pa kuyeretsedwa kumodzi kwa zitsanzo zoyendetsedwa.Pazitsanzo za zomera za polyphenol polysaccharides, tikupangira kuti muyese Plant Total RNA Isolation Kit Plus.
4. Kutentha kwa centrifuge ndikotsika kwambiri.
Kudzipatula kwathunthu kwa RNA ndi kuyeretsedwa, kupatula kusokonezeka kwa nayitrogeni wamadzimadzi amtundu wa zitsanzo, masitepe onse amachitikira kutentha (20-25 ° C).Ma centrifuge ena otsika amakhala ndi kutentha kosachepera 20 °C, zomwe zingayambitse kutsekeka kwa DNA-Cleaning Column ndi/kapena RNA-Oly Column.Izi zikachitika, ikani kutentha kwa centrifuge ku 20-25 °C ndikutenthetsa chisakanizo cha lysis ndi/kapena kuwonjezera kwa ethanol kupatukana kwamphamvu kwambiri mpaka 37 °C.
Palibe RNA yotengedwa kapena zokolola za RNA ndizochepa
Nthawi zambiri pamakhala zinthu zambiri zomwe zimakhudza kuchira bwino, monga: zitsanzo za RNA, njira yogwirira ntchito, kuchuluka kwa elution, ndi zina zambiri.
Kusanthula kwazomwe zimayambitsa zomwe zili pansipa:
1.Kusambira kwa ayezi kapena kutentha kochepa (4 ° C) centrifugation kunachitika panthawi ya opaleshoni.
Malangizo: Gwiritsani ntchito kutentha kwapakati (15-25 ° C) panthawi yonseyi, musamasambitse madzi oundana komanso kutentha kwapakati.
2.RNA yawonongeka chifukwa cha kusungidwa kosayenera kwa chitsanzo kapena kusungidwa kwa nthawi yaitali kwa chitsanzo.
Malangizo: Zitsanzo zomwe zangotengedwa kumene ziyenera kusungidwa mwachangu mu nayitrogeni wamadzimadzi, kenako kusungidwa pa -80 ° C kwa nthawi yayitali, kupewa kuzizira mobwerezabwereza ndi kusungunuka kwa zitsanzo;kapena nthawi yomweyo zilowerereni zitsanzo mu RNA stabilizer RNAlater solution (zitsanzo za nyama).
3.Kugawika kwachitsanzo kosakwanira ndi lysis kumabweretsa kutsekeka kwa gawo loyeretsa.
Lingaliro: Mukamagaya minofu, chonde onetsetsani kuti minofuyo ili pansi mokwanira, ndipo mwamsanga mutumize ku Buffer PSL1 yokonzekera kale (tsimikizirani kuti gawo lolondola la β-ME lawonjezeredwa, onani sitepe 1 ya ndondomekoyi).
4.Chidziwitso chinawonjezedwa molakwika.
Yesani: Onetsetsani kuti RNase-Free ddH2O amadonthozedwa mkatikati mwa nembanemba yoyeretsa.
5.Voliyumu yolondola ya ethanol mtheradi sanawonjezedwe ku Buffer PSL2 kapena Buffer PRW2.
Malingaliro: Chonde tsatirani malangizo, onjezani voliyumu yolondola ya ethanol ku Buffer PSL2 ndi Buffer PRW2 ndipo sakanizani bwino zida zisanagwiritsidwe.
6.Kuchuluka kwa chitsanzo cha minofu ndi kosayenera.
Malingaliro: Gwiritsani ntchito 50 mg ya minofu pa 500 μl ya Buffer PSL1.Kugwiritsa ntchito minofu yambiri kumachepetsa kuchuluka kwa RNA yotulutsidwa ndipo chiyero cha RNA chotsatira chidzachepetsedwanso.Tikukulimbikitsani kuti mlingo woyambirira wa chitsanzo usapitirire 50 mg pa ntchito yochotsa RNA.
7. Voliyumu yofotokozera molakwika kapena kusakwanira.
Lingaliro: Voliyumu yosadziwika ya gawo loyeretsera ndi 50-200 μl;ngati mphamvu ya elution siili yokhutiritsa, tikulimbikitsidwa kuwonjezera nthawi kutentha kwapakati mutatha kuwonjezera RNase-Free ddH2O, monga 5-10min.
8.Mzere woyeretsedwa uli ndi zotsalira za ethanol mutatsuka ndi Buffer PRW2.
Lingaliro: Ngati chubu chopanda kanthu chili ndi centrifuged kwa 1 min ndipo pali ethanol yotsalira mutatsuka mu Buffer PRW2, mukhoza kuwonjezera nthawi ya chubu chopanda kanthu centrifugation ku 2 min, kapena ikani ndime yoyeretsera kutentha kwa mphindi 5 kuti muchotse ethanol yotsalira.
9.Chidacho chinagwiritsidwa ntchito molakwika.
Malingaliro: Pazitsanzo za mbewu za polyphenolic polysaccharides, kugwiritsa ntchito zida wamba monga Plant Total RNA Isolation Kit sangathe kupeza zitsanzo zabwino za RNA.Tikukulangizani kuti mugwiritse ntchito Plant Total RNA IsolationKit Plus, yomwe idapangidwira mwapadera zitsanzo za mbewu za polyphenolic polysaccharide.Chida chopangidwa mwapadera chochotsera RNA kuchokera ku zitsanzo za mbewu za polyphenol ndi polysaccharide.
Mtengo wa OD260/OD280 ndiwotsika
Kusintha kwa RNA ndi ddH2O ndi kugwiritsidwa ntchito powerengera ma spectrophotometer kumapangitsa kuti OD260/OD280 ikhale yotsika.Timalimbikitsa kugwiritsa ntchito 10 mM Tris-HCl, pH 7.5 (m'malo mwa RNase-Free ddH2O to elute RNA) kuti mupeze mfundo zolondola za OD260/OD280, onani “RNA Concentration and Purification Assays” patsamba 19.
RNA yoyeretsedwa ndiyonyozeka
Ubwino wa RNA yoyeretsedwa umagwirizana ndi zinthu monga kusungidwa kwa zitsanzo, kuipitsidwa kwa RNase, ndi kusintha.
Kusanthula kwazomwe zimayambitsa:
Zitsanzo za 1.Tissue sizinasungidwe mu nthawi pambuyo pa kusonkhanitsa.
Malangizo: Ngati zitsanzo za minofu sizikugwiritsidwa ntchito pakapita nthawi, chonde zisungeni mu nayitrogeni wamadzimadzi pa kutentha kochepa kapena kuwasamutsira ku -80 ° C kuti asungidwe kwa nthawi yaitali mutatha kuzizira mofulumira mu nayitrogeni wamadzimadzi, kapena kumiza mwamsanga zitsanzo mu RNA stabilizer RNAlater solution (zitsanzo za nyama).Pochotsa RNA, yesani kugwiritsa ntchito zitsanzo za minofu zomwe zasonkhanitsidwa mwatsopano.
2.Kuzizira kobwerezabwereza ndi kusungunuka kwa zitsanzo za minofu.
Yesani izi: Posunga zitsanzo za minofu, ndi bwino kuziduladula kuti zisungidwe, ndi kuchotsamo mbali ina yake pozigwiritsira ntchito kupeŵa kuwonongeka kwa RNA kochititsidwa ndi kuzizira kobwerezabwereza ndi kusungunuka kwa zitsanzozo.
3.RNase imayambitsidwa mchipinda chogwirira ntchito kapena osavala magolovesi otaya, masks, ndi zina.
Lingaliro: Kuyesa kwa RNA m'zigawo kumachitidwa bwino pamachitidwe osiyanasiyana a RNA, ndipo tebulo la labotale liyenera kutsukidwa kuyesako kusanachitike, ndipo magolovesi otayika ndi masks ayenera kuvalidwa panthawi yoyesera kuti apewe kuwonongeka kwa RNA komwe kumachitika chifukwa cha kukhazikitsidwa kwa RNase kwambiri.
4.The reagent imakhudzidwa ndi RNase panthawi yogwiritsira ntchito.
Yesani: M'malo mwake lowetsani zida zatsopano za RNA zotulutsira mbewu pazoyeserera zofananira.
5.Machubu a centrifuge ndi malangizo a pipette omwe amagwiritsidwa ntchito pakusintha kwa RNA ali ndi kachilombo ka RNase.
Malangizo: Onetsetsani kuti machubu a centrifuge, nsonga za pipette, pipettes, ndi zina zotero zomwe zimagwiritsidwa ntchito pochotsa RNA zonse ndi RNase-Free.
Mabuku a Malangizo:
