Malangizo owunikira mavuto

The following is an analysis of the problems that might be encountered in the extraction of viral RNA. We wish it would be helpful to your experiment. In addition, for other experimental or technical problems other than operating instructions and problem analysis, we have dedicated technical support to help you. Contact us if you need at : 028-83361257or E-mail：Tech@foregene.com。

Palibe RNA yomwe ingatulutsidwe kapena zokolola za nucleic acid ndizochepa

Nthawi zambiri pamakhala zinthu zambiri zomwe zimakhudza kuchira bwino, monga: zitsanzo za RNA, njira yogwirira ntchito, voliyumu, etc..

Kusanthula kwazomwe zimayambitsa:

1.Ice kusamba kapena otsika kutentha (4 ° C) centrifugation pa ntchito.

Malangizo: Kutentha kwachipinda (15-25 ° C), osasamba madzi oundana komanso kutentha kwa centrifuge.

2. Kusungidwa kwachitsanzo kosayenera kapena kusungirako zitsanzo kwa nthawi yayitali.

Malangizo: Sungani zitsanzo pa -80 ° C kapena kuzizira mu nayitrogeni wamadzimadzi, ndipo pewani kugwiritsa ntchito kuzizira kobwerezabwereza;yesani kugwiritsa ntchito zitsanzo zomwe zasonkhanitsidwa mwatsopano kuti muchotse RNA.

3.Kusakwanira kwachitsanzo lysis

Malangizo: Chonde onetsetsani kuti zitsanzo ndi njira yogwirira ntchito (Linear Acrylamide) yasakanizidwa bwino ndikuyimitsidwa kwa mphindi 10 kutentha (15-25 ° C)

4.Chidziwitso chinawonjezedwa molakwika

Malangizo: Onetsetsani kuti RNase-Free ddH2O yawonjezedwa pakati pa nembanemba ya gawo loyeretsa.

5.Voliyumu yolakwika ya ethanol ya anhydrous mu Buffer viRW2

Yesani: Chonde tsatirani malangizo, onjezani voliyumu yolondola ya ethanol ya anhydrous ku Buffer viRW2 ndikusakaniza bwino musanagwiritse ntchito zida.

6.Kugwiritsira ntchito chitsanzo molakwika.

Malingaliro: 200µl ya zitsanzo pa 500μl ya Buffer viRL.Kuchuluka kwachitsanzo kumabweretsa kuchepa kwa RNA m'zigawo.

7. Voliyumu yolakwika kapena kusakwanira.

Lingaliro: Voliyumu yosawoneka bwino ya gawo loyeretsera ndi 30-50μl;ngati mphamvu ya elution siyikukhutiritsa, tikulimbikitsidwa kuwonjezera RNase-Free ddH yotenthetserapo.₂O ndikuwonjezera nthawi yoyika kutentha, monga 5-10min

8.Kuyeretsa kumakhala ndi zotsalira za ethanol mutatsuka mu Buffer viRW2.

Lingaliro: Ngati Mowa utsalirabe mutatha kutsuka mu Buffer viRW2 ndi centrifugation yopanda kanthu-chubu kwa 2min, ndime yoyeretsera imatha kusiyidwa kutentha kwa mphindi 5 pambuyo pa chubu chopanda kanthu kuti muchotse ethanol yotsalira.

Kuwonongeka kwa mamolekyu oyeretsedwa a RNA

Ubwino wa RNA yoyeretsedwa umagwirizana ndi zinthu monga kusungirako zitsanzo, kuipitsidwa kwa RNase, ndi ntchito.

Kusanthula kwazomwe zimayambitsa:

1.Zitsanzo zosonkhanitsidwa sizinasungidwe mu nthawi.

Yesani: Ngati chitsanzocho sichigwiritsidwa ntchito pakapita nthawi, chonde sungani pa -80 ℃ kapena nayitrogeni wamadzimadzi nthawi yomweyo.Pochotsa mamolekyu a RNA, yesani kugwiritsa ntchito zitsanzo zomwe zasonkhanitsidwa mwatsopano ngati kuli kotheka.

2.Zitsanzo zosonkhanitsa zinali kuzizira ndi kusungunuka mobwerezabwereza.

Yesani: Pewani kuzizira kobwerezabwereza ndi kusungunuka (osapitirira kamodzi) panthawi yosonkhanitsa ndi kusunga, apo ayi zokolola za nucleic acid zidzachepa.

3.RNase idayambitsidwa m'chipinda chopangira opaleshoni kapena palibe magolovesi otaya, masks, ndi zina zambiri.

Lingaliro: Kutulutsa kwa kuyesa kwa mamolekyu a RNA kumachitidwa bwino m'chipinda chapadera cha RNA, ndipo tebulo loyesera limatsukidwa kuyesako kusanachitike.Valani magolovesi ndi masks otayika panthawi yoyesera kuti mupewe kuwonongeka kwa RNA komwe kumachitika chifukwa cha kuyambika kwa RNase.

4.The reagent imakhudzidwa ndi RNase panthawi yogwiritsira ntchito.

Yesani: Bwezerani ndi Viral RNA Isolation Kit yatsopano pazoyeserera zofananira.

5.Kuwonongeka kwa RNase kwa machubu a centrifuge, nsonga za pipette, ndi zina zotero.

Mamolekyu oyeretsedwa a RNA adakhudza zoyeserera zakutsika

Mamolekyu a RNA oyeretsedwa ndi gawo loyeretsera adzakhudza kuyesa kumtunda ngati pali ma ion amchere ambiri kapena mapuloteni, monga: reverse transcript, Northern Blot, etc..

1.Pali ma ion amchere otsala mu ma molekyulu a RNA opangidwa.

Malangizo: Onetsetsani kuti voliyumu yolondola ya ethanol ya anhydrous yawonjezedwa ku Buffer viRW2, ndikutsuka ndime yoyeretsa kawiri molingana ndi liwiro lolondola la centrifugation pamalangizo opangira; Ngati pali ma ion amchere otsala, mutha kuwonjezera Buffer viRW2 pagawo loyeretsera, ndikuyisiya kutentha kwa 5min.Ndiye kuchita centrifugation kuchotsa mchere ayoni kuipitsidwa kwambiri

2.Pali ethanol yotsala mu mamolekyu a RNA opangidwa

Lingaliro: mutatsimikizira kuti mizati yoyeretsera yatsukidwa ndi Buffer viRW2, chitani chopanda kanthu-chubu centrifugation malinga ndi liwiro la centrifugal pa malangizo ogwiritsira ntchito.Ngati pali Mowa otsala, akhoza kusiyidwa kwa mphindi 5 kutentha firiji pambuyo chopanda kanthu chubu centrifugation kuchotsa Mowa otsala kumlingo waukulu.

Mabuku a Malangizo:

Viral RNA Isolation Kit Instruction Manual