Maginito Matishu / Cell / Magazi Onse RNA Kit
Mawonekedwe
■ Mosavuta komanso mwachangu: RNA yathunthu imatha kupezeka mkati mwa mphindi 60.
■ Kutulutsa kwakukulu: Ikhoza kukwaniritsa zofunikira pakupanga pamanja komanso kutulutsa batch pama pulatifomu ena apamwamba.
■ Otetezeka komanso osakhala ndi poizoni: Palibe reagent monga phenol / chloroform yofunikira.
Mfundo
Mtundu: Maginito amtundu wamtundu wamtundu.
Chitsanzo: Minofu, maselo ndi magazi.
Cholowera: RNA Yonse
Kuyambira voliyumu: 5-20 mg, osapitilira 1 × 107, 0,1-1.5 ml
Nthawi yogwira ntchito: 60 min
Ntchito zotsikira: RT-PCR / RT-qPCR, NGS library library, ndi zina zambiri.
Zogulitsa zonse zimatha kusinthidwa kukhala ODM / OEM. Zambiri,chonde dinani Makonda Service (ODM / OEM)
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RNA yonse idatengedwa kuchokera ku chiwindi cha 20 mg rat ndi TIANGEN Magnetic Tissue / Cell / Blood Yonse RNA Kit ndi zinthu zina zofunikira kuchokera kwa omwe amapereka. 5 μl ya 200 μl eluate idakwezedwa 1% agarose gel electrophoresis, 6 V / cm. Kutsiliza: Kuchulukitsa, kuyera ndi kukhazikika kwa TIANGEN Maginito Tishu / Cell / Magazi Onse RNA Kit ndiabwino kuposa ena ogulitsa. |
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RNA yonse idatengedwa kuchokera ku 20 mg ya impso za makoswe ndi TIANGEN Magnetic Tissue / Cell / Blood Yonse RNA Kit ku TIANGEN TGuide S32 kapena Thermo Kingfisher Flex96 motsatana. 5 μl ya 200 μl eluate idakwezedwa 1% agarose gel electrophoresis, 6 V / cm. Cholemba: TIANGEN D15000 Chikhomo cha DNA. Kutsiliza: Maginito Tissue / Cell / Blood Yonse RNA Kit ili ndi zokolola zabwino komanso zoyera muma extractor osiyanasiyana. |
A-1 Cell lysis kapena homogenization sikokwanira
---- Kuchepetsa ntchito nyemba, kuonjezera kuchuluka kwa gawo lotetezera lysis, kuonjezera homogenization ndi nthawi lysis.
A-2 Zitsanzo kuchuluka ndi waukulu kwambiri
---- Kuchepetsa kuchuluka kwa nyemba zomwe mukugwiritsa ntchito kapena kuonjezera kuchuluka kwa lysis buffer.
A-1 Kusakwanira kwa cell lysis kapena homogenization
---- Kuchepetsa ntchito nyemba, kuonjezera kuchuluka kwa gawo lotetezera lysis, kuonjezera homogenization ndi nthawi lysis.
A-2 Zitsanzo kuchuluka ndi waukulu kwambiri
- Chonde onani kuthekera kwakukulu kokonza.
A-3 RNA siyosankhidwa kwathunthu kuchokera pagawo
---- Mukatha kuwonjezera madzi opanda RNase, siyani kwa mphindi zochepa centrifuging.
A-4 Ethanol mumlengalenga
- Pambuyo kutsukidwa, centrifuge kachiwiri ndikuchotsa chotsuka chotsuka momwe zingathere.
A-5 Cell chikhalidwe sing'anga sichichotsedweratu
- Mukamasonkhanitsa maselo, chonde onetsetsani kuti muchotse sing'anga momwe mungathere.
A-6 Maselo osungidwa mu RNAstore sakhala otsekemera kwenikweni
---- RNAstore osalimba ndi wamkulu kuposa sing'anga wamba wamaselo; kotero mphamvu ya centrifugal iyenera kukulitsidwa. Akuti centrifuge pa 3000x g.
Zolemba za A-7 Low RNA komanso kuchuluka kwake mchitsanzo
- Gwiritsani ntchito chitsanzo chabwino kuti muwone ngati zokolola zochepa zimayambitsidwa ndi chitsanzocho.
A-1 Nkhaniyi siyatsopano
- Minyewa yatsopano imayenera kusungidwa mu nayitrogeni wamadzi nthawi yomweyo kapena kuyiyika mu reagent ya RNAstore kuti zitsimikizidwe kuti zimachokera.
A-2 Zitsanzo kuchuluka ndi waukulu kwambiri
---- Kuchepetsa nyemba kuchuluka.
A-3 RNase kuipitsidwan
- Ngakhale gawo lotetezedwa mulibe RNase, ndikosavuta kuipitsa RNase panthawi yazochulukirapo ndipo iyenera kusamalidwa.
Kuwononga kwa A-4 Electrophoresis
---- Sinthanitsani cholumikizira cha electrophoresis ndikuwonetsetsa kuti zofunikira ndi Kutsegula Buffer zilibe vuto la RNase.
A-5 Kutsitsa kwambiri kwa electrophoresis
---- Kuchepetsa kuchuluka kwa potsegula, kutsitsa kwa chitsime chilichonse sikuyenera kupitirira 2 μg.
A-1 Zitsanzo kuchuluka ndi waukulu kwambiri
---- Kuchepetsa nyemba kuchuluka.
A-2 Zitsanzo zina zimakhala ndi DNA yambiri ndipo amatha kuchiritsidwa ndi DNase.
- Chitani chithandizo cha RNase-Free DNase ku yankho la RNA, ndipo RNA itha kugwiritsidwa ntchito mwachindunji poyeserera pambuyo pothandizidwa, kapena itha kuyeretsedwa ndi zida zoyeserera za RNA.
Zamagalasi, zophikidwa pa 150 ° C kwa 4 h. Pazitsulo zapulasitiki, zomizidwa mu 0,5 M NaOH kwa mphindi 10, kenako kutsukidwa bwino ndi madzi opanda RNase kenako ndikuwotcha kuti muchotse RNase. Ma reagents kapena mayankho omwe amagwiritsidwa ntchito poyeserera, makamaka madzi, ayenera kukhala opanda RNase. Gwiritsani ntchito madzi opanda RNase pamakonzedwe onse a reagent (onjezerani madzi mu botolo loyera lagalasi, onjezerani DEPC pamlingo womaliza wa 0,1% (V / V), gwedezani usiku umodzi ndi autoclave).
Zamgululi siyana
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