FAQ 's noʻO ka pahu hoʻokaʻawale RNA holoʻokoʻa holoholona

ʻO kēia ka nānā ʻana o nā pilikia āu e hālāwai ai makaʻiʻo holoholona/cell RNA 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.

ʻAʻole lawe ʻia ka RNA a i ʻole nā ​​​​hua RNA

Nui nā ʻano like ʻole e pili ana i ka pono o ka hoʻihoʻi hou ʻana, e like me: kiko kiko RNA maʻiʻo, ke ʻano o ka hana, ka nui elution, etc.

1. Hana ʻia ka ʻauʻau hau a i ʻole cryogenic (4 °C) centrifugation i ka wā o ka hana.

Manaʻo: E hana ma ka lumi wela (15-25 ° C) i ke kaʻina holoʻokoʻa, mai ʻauʻau hau a centrifuge i nā haʻahaʻa haʻahaʻa.

2. Ka mālama pono ʻole ʻana i ka laʻana a i ʻole ka nui o ka manawa e mālama ai i ka laʻana.

Manaʻo Manaʻo: E kūʻai i nā laʻana ma -80 °C a i ʻole i loko o ka hau hau i loko o ka wai hau a pale i ka hoʻohana pinepine ʻana i ka hau hau;e ho'āʻo e hoʻohana i ka ʻiʻo hou a i ʻole nā ​​​​pūnaewele moʻomeheu no ka unuhi ʻana i ka RNA.

3. lawa ole la'ana lysis.

Manaʻo Manaʻo: I ka wā e hoʻohui ai i ka ʻiʻo, e hōʻoia i ka hoʻomaʻemaʻe ʻia ʻana o ka ʻiʻo a ua hoʻokaʻawale ʻia nā mea kikoo e wehewehe i ka hoʻokuʻu ʻana o RNA.

4. ʻAʻole i hoʻohui pono ʻia ka eluent.

Manaʻo: E hōʻoia i ka RNase-Free ddH₂Hoʻohui ʻia ʻo O i kahi dropwise i ka waena o ka membrane kolamu hoʻomaʻemaʻe.

5. ʻAʻole i hoʻohui ʻia ka leo kūpono o ka ethanol piha i ka Buffer RL2 a i ʻole Buffer RW2.

Manaʻo Manaʻo: E hahai i nā kuhikuhi, e hoʻohui i ka leo kūpono o ka ethanol piha i ka Buffer RL2 a me Buffer RW2 a hui maikaʻi ma mua o ka hoʻohana ʻana i ka pahu.

6. ʻAʻole kūpono ke ʻano o ka laʻana.

Manaʻo: E hoʻohana i ka 10-20 mg o ka ʻiʻo a i ʻole (1-5) × 10⁶nā pūnaewele no 500 μl buffer RL1, no ka mea, hiki i ka hoʻohana ʻana i ka ʻiʻo nui ke hopena i ka hōʻemi ʻana i ka RNA extraction.

7. Ka nui elution kūpono ʻole a i ʻole ka elution piha ʻole.

Manaʻo: ʻO ka nui elution o ke kolamu hoʻomaʻemaʻe ʻo 50-200 μl;inā ʻaʻole ʻoluʻolu ka hopena elution, pono ia e hoʻolōʻihi i ka manawa kau wela o ka lumi ma hope o ka hoʻohui ʻana i ka RNase-Free ddH preheated.₂ʻO, e laʻa no 5-10 min.

8. He koena ethanol ka kolamu hoʻomaʻemaʻe ma hope o ka holoi ʻana o Buffer RW2.

Manaʻo Manaʻo: Inā loaʻa ke koena ethanol ma hope o ka holoi ʻana o Buffer RW2, ʻo ka centrifugation paipu ʻole no 1min, hiki ke hoʻonui ʻia ka manawa no ka hana centrifugation pahu ʻole i 2min, a i ʻole ke kolamu hoʻomaʻemaʻe hiki ke kau ʻia ma ka lumi wela no 5 min e wehe pono i ke koena ethanol.

Hoʻohaʻahaʻa ʻia ka RNA hoʻomaʻemaʻe

ʻO ka maikaʻi o ka RNA i hoʻomaʻemaʻe ʻia e pili ana i nā mea e like me ka mālama ʻana i ka laʻana, RNase contamination, a me ka manipulation, etc.

1. ʻAʻole mālama ʻia nā ʻāpana kiko i ka manawa.

Manaʻo Manaʻo: Inā ʻaʻole hoʻohana ʻia nā laʻana a i ʻole nā ​​​​pūnaewele i ka manawa kūpono ma hope o ka hōʻiliʻili ʻana, e mālama koke i ka cryopreserve ma -80 °C a i ʻole ka wai nitrogen.No ka unuhi ʻana i ka RNA, e hoʻohana i kahi ʻiʻo i lawe ʻia a i ʻole ka laʻana cell i ka wā hiki.

2. ʻO ka hoʻoheheʻe hou ʻia ʻana o nā laʻana kiko.

Manaʻo Manaʻo: I ka wā e mālama ai i nā laʻana ʻiʻo, ʻoi aku ka maikaʻi e ʻoki ʻia i mau ʻāpana liʻiliʻi no ka mālama ʻana, a e wehe i kekahi o nā ʻāpana i ka wā e hoʻohana ai ia mau mea e pale aku ai i ka hoʻoheheʻe pinepine ʻana o ka hāpana a me ka hōʻino ʻana o RNA.

3. Hoʻokomo ʻia ʻo RNase a ʻaʻole hoʻohana i nā mīkina lima hoʻolele, masks, etc. i ka wā o ka hana.

Manaʻo Manaʻo: Hana maikaʻi ʻia nā hoʻokolohua RNA extraction ma nā keʻena hoʻoponopono RNA kaʻawale a holoi ʻia ka papaʻaina ma mua o ka hoʻokolohua.

E hoʻohana i nā mīkina lima a me nā maka i ka wā o ka hoʻokolohua e hōʻemi i ka hōʻemi ʻana o RNA ma muli o ka hoʻokomo ʻana o RNase.

4. Ho'ohaumia 'ia nā reagents me RNase i ka wā e ho'ohana ai.

Manaʻo Manaʻo: E hoʻololi me kahi Animal Total RNA Isolation Kit no nā hoʻokolohua pili.

5. ʻO nā paipu centrifuge, nā ʻōlelo aʻoaʻo, a me nā mea ʻē aʻe i hoʻohana ʻia i ka RNA manipulation ua haumia me RNase.

Manaʻo Manaʻo: E hōʻoia i ka RNase-Free nā paipu centrifuge, nā ʻōlelo aʻoaʻo, nā pipette, a me nā mea ʻē aʻe i hoʻohana ʻia i ka unuhi RNA.

Hoʻopili ka RNA i hoʻomaʻemaʻe ʻia i nā hoʻokolohua ma lalo

Hoʻomaʻemaʻe ʻia ʻo RNA e ke kolamu hoʻomaʻemaʻe, inā ʻoi aku ka nui o nā ion paʻakai, ʻoi aku ka nui o ka protein ma lalo o ka hoʻokolohua, e like me: reverse transcription, Northern Blot et al.

1. He koena iona paakai ko ka RNA i hooleiia.

Manaʻo Manaʻo: E hōʻoia ua hoʻohui ʻia ka nui kūpono o ka ethanol i ka Buffer RW2 a hana i 2 holoi kolamu hoʻomaʻemaʻe ma ka wikiwiki centrifugal i hōʻike ʻia no ka hana;inā loaʻa kekahi koena ion paʻakai, waiho i ke kolamu hoʻomaʻemaʻe iā Buffer RW2 no 5 min ma ka lumi wela a hana centrifugation e hoʻonui i ka wehe ʻana o ka paʻakai.

2. Ke koena ethanol i loko o ka RNA i hoʻoheheʻe ʻia.

Manaʻo Manaʻo: E hōʻoia ma hope o ka holoi ʻana i ka RW2, e hoʻokō i ka hana centrifugation paipu ʻole ma ka wikiwiki centrifugation i hōʻike ʻia no ka hana, e hoʻonui i ka manawa o ka hana centrifugation paipu ʻole i 2 min inā he koena ethanol, a i ʻole waiho ʻia ma ka lumi wela no 5 min ma hope o ka centrifugation pahu hakahaka e hoʻonui i ka wehe ʻana o ke koena ethanol.

ʻO ka hoʻokaʻawale ʻana i nā ʻano laʻana nulceic acid

Hiki i nā pahu hoʻokaʻawale RNA o Foregene ke loaʻa i ka hoʻokaʻawale ʻana a me ka unuhi ʻana o ka RNA mai nā kumu hoʻohālike aʻe: ʻiʻo holoholona, ​​mea kanu, cell, viral, blood, etc.

Pehea wau e mālama ai i ka pahu

ʻO ka mālama ʻana i ka lumi wela no 24 mahina.