tsatirani mgwirizano”, zikugwirizana ndi zomwe msika ukufunikira, kujowina pampikisano wamsika ndi mtundu wake wabwino momwemonso momwe zimaperekera chithandizo chokwanira komanso chapamwamba kwa makasitomala kuti awalole kukhala opambana kwambiri.Kutsata kampaniyo, ndikosangalatsa kwamakasitomala a China New Product Best Selling Viral DNA&Rna Extraction Kit, Takulitsa bizinesi yathu ku Germany, Turkey, Canada, USA, Indonesia, India, Nigeria, Brazil ndi zigawo zina zapadziko lonse lapansi.Tikuyesetsa kukhala m'modzi mwa ogulitsa bwino kwambiri padziko lonse lapansi.
tsatirani mgwirizano”, zikugwirizana ndi zomwe msika ukufunikira, kujowina pampikisano wamsika ndi mtundu wake wabwino momwemonso momwe zimaperekera chithandizo chokwanira komanso chapamwamba kwa makasitomala kuti awalole kukhala opambana kwambiri.Kutsata kwa kampaniyo, ndikosangalatsa kwamakasitomalaChina Rna&DNA Extraction Kit ndi Nucleic Acid Extraction, Tikuyembekezera kukhazikitsa ubale wopindulitsa ndi inu potengera zinthu zathu zapamwamba komanso zothetsera, mitengo yabwino komanso ntchito zabwino kwambiri.Tikukhulupirira kuti mankhwala athu adzakubweretserani chokumana nacho chokoma ndi kunyamula kumverera kukongola.
Mabuku a Malangizo:Bzalani Total RNA Isolation Kit Plus Instruction Manual

tsatirani mgwirizano”, zikugwirizana ndi zomwe msika ukufunikira, kujowina pampikisano wamsika ndi mtundu wake wabwino momwemonso momwe zimaperekera chithandizo chokwanira komanso chapamwamba kwa makasitomala kuti awalole kukhala opambana kwambiri.Kutsata kampaniyo, ndikosangalatsa kwamakasitomala a China New Product Best Selling Viral DNA&Rna Extraction Kit, Takulitsa bizinesi yathu ku Germany, Turkey, Canada, USA, Indonesia, India, Nigeria, Brazil ndi zigawo zina zapadziko lonse lapansi.Tikuyesetsa kukhala m'modzi mwa ogulitsa bwino kwambiri padziko lonse lapansi.
China New Product China Rna&DNA Extraction Kit ndi Nucleic Acid Extraction, Tikuyembekezera kukhazikitsa ubale wopindulitsa ndi inu potengera zinthu zathu zapamwamba komanso zothetsera, mitengo yabwino komanso ntchito zabwino kwambiri.Tikukhulupirira kuti mankhwala athu adzakubweretserani chokumana nacho chokoma ndi kunyamula kumverera kukongola.

Chigawo chalumikizidwa

Pambuyo potsekedwa ndime, zokolola za RNA zimachepetsedwa kapena zosatheka kuyeretsa RNA, ndipo kuchuluka kwa RNA komwe kumapezeka kumakhala kochepa.

Kusanthula zomwe zimachitika kawirikawiri:

1. Zitsanzo zopuma sizili bwino.

Kusweka kwachitsanzo sikumapangitsa kuti DNA-CLEANING COLUMN itsekedwe, pomwe imakhudza zokolola za RNA ndi mtundu wake.Mpofunika mofulumira akupera ntchito okwanira madzi asafe pamene inu anaswa zitsanzo, Yesani kuphwanya chitsanzo selo khoma, selo nembanemba ndi minofu ina.Pazitsanzo za mbewu za polyol polysaccharides, timalimbikitsa kuti mugwiritse ntchito Plant Total RNA IsolATION KIT PLUS.

2. Mukayamwa chopatukana champhamvu champhamvu ndi DNA-Cleaning Column, ma cell omwe angagawike amatha kupumira.

Maselo ogawanika a cell omwe atengedwa adzapangitsa RNA-ONLY Column yomwe idzatsekerezedwa pamene RNA adsorption operation ikuchitika (onani sitepe 6).Tikukulangizani mosamala mukayamwa chodabwitsa ichi kuti mupewe kuyamwa zinyalala zama cell.

3. Zitsanzo za ndalama zoyambira ndizochuluka kwambiri.

Kugwiritsa ntchito kwambiri zitsanzo kumabweretsa kugawika kwachitsanzo kosakwanira kapena kusakwanira kwa cell lysis ndi Buffer PSL1, zomwe zimabweretsa kutsekeka kwa gawo loyeretsa pakuyeretsedwa.Plant Total RNA Isolation Kit Chitsanzo chilichonse choyeretsedwa ndi 50 mg.Pazitsanzo za mbewu za polyol polysaccharides, tikupangira kuti muyese Plant Total RNA IsolATION KIT PLUS.

4. Kutentha kwa centrifuge ndikotsika kwambiri.

Njira yonse yodzipatula ndi kuyeretsa RNA imachitika kutentha (20-25).°C), kupatula kuti minofu yachitsanzo imasweka ndi nayitrogeni wamadzimadzi. Kutentha kwa ma cryogenic centrifuges ndi otsika kuposa 20.℃, zomwe zingayambitse kutsekeka kwa DNA-Cleaning Column ndi/kapena RNA-Only Column.Izi zikachitika, ikani kutentha kwa centrifuge ku 20-25℃,ndionetsetsani kuti lysis osakaniza ndi/kapena Mowa-anawonjezera supernatant anali preheated kuti 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.

Yesani: Gwiritsani ntchito kutentha kwa chipinda (15-25°C) mu ndondomeko yonse, musachite ayezi kusamba ndi otsika kutentha centrifugation.

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.Kusakwanira kwachitsanzo kugawikana ndi lysis kumayambitsa 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 yadonthezedwa pakati pa nembanemba yazakudya.

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 yosadziwika bwino kapena kutulutsa kosakwanira.

Lingaliro: Voliyumu yosadziwika ya gawo loyeretsera ndi 50-200 μl;ngati elution zotsatira si zogwira mtima, Ndi bwino kuwonjezera nthawi firiji pambuyo kuwonjezera preheated RNase-Free ddH2O, monga 5-10min.

8.Mzere woyeretsa uli ndi zotsalira za ethanol mutatsuka ndi BufferPRW2.

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

RNA elution yokhala ndi ddH2O komanso yogwiritsidwa ntchito powerengera ma spectrophotometer imapangitsa 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:

Bzalani Total RNA Isolation Kit Plus Instruction Manual