Ke alakaʻi hoʻoponopono pilikia

The following is an analysis of the problems that may be encountered in the extraction of plant genomic DNA, hoping to be helpful to your experiments. In addition, for other experimental or technical problems other than operation instructions and problem analysis, we have dedicated technical support to help you. If you have any needs, please contact us: 028-83360257 or E-mali: Tech@foregene.com.

Haʻahaʻa haʻahaʻa a ʻaʻohe DNA

Nui nā kumu e pili ana i ka hua o ka DNA genomic, me ke kumu o ka hāpana, ka makahiki o ka hāpana, nā kūlana mālama o ka hāpana, a me ka hana.

ʻAʻole hiki ke loaʻa ka DNA Genomic i ka wā o ka unuhi ʻana

1. Hoʻopaʻa pono ʻole ʻia a mālama ʻia paha nā mea kikoo no ka lōʻihi loa, e hopena i ka hōʻino ʻana o ka DNA genomic.

Manaʻo: E mālama i nā laʻana o ka ʻiʻo i loko o ka nitrogen wai a i ʻole -20°C;e ho'āʻo e hoʻohana i nā laʻana hou i hōʻiliʻili ʻia no ka unuhi ʻana i ka DNA genomic.

2. ʻAʻole hiki ke unuhi ʻia ka DNA genomic pili i ka liʻiliʻi loa.

Manaʻo Manaʻo: No nā laʻana ʻiʻo i mālama ʻia no ka manawa lōʻihi a i ʻole ka hoʻohaʻahaʻa ʻana i ka DNA genomic koʻikoʻi, hiki ke hoʻonui ʻia ka nui o nā laʻana iʻa no ka unuhi ʻana i ka DNA genomic nui.Hiki ke hoʻoholo ʻia ka nui o ka hāpana e like me ka pono o ka DNA, akā ʻaʻole pono ka laʻana hou ma mua o 100mg, a ʻaʻole pono ka hāpana maloʻo ma mua o 30mg.

3. ʻAʻole i lepo ka hāpana me ka naikokene wai a waiho ʻia no ka lōʻihi loa ma hope o ka wai nitrogen.

Manaʻo Manaʻo: I ka wā o ka unuhi ʻana i ka DNA, pono e hoʻoheheʻe ʻia ka hāpana me ka nitrogen wai e wāwahi ai i ka paia o ke kelepona;ma hope o ka wili ʻana, e ʻoluʻolu e hoʻololi i ka pauka hāpana i PL1 i hoʻomehana ʻia ma 65°C i ka hikiwawe loa (ke hoʻoheheʻe ʻia ka lepo lepo, e hoʻomaka ana ka DNA genomic e hoʻohaʻahaʻa koke) .

4. ʻO ka mālama pono ʻole ʻana o Foregene Protease ka hopena i ka hoʻemi ʻana a i ʻole ka hana ʻole.

Manaʻo: E hōʻoia i nā kūlana mālama o Foregene Protease a i ʻole e hoʻololi iā ia me kahi Foregene Protease hou no ka hydrolysis enzymatic.

5. Ua mālama pono ʻole ʻia a mālama ʻia paha ka pahu no ka lōʻihi loa, e hāʻule ana kekahi mau mea i loko o ka pahu.

Manaʻo Manaʻo: E kūʻai mai i kahi pahu hoʻoheheʻe DNA genomic mea kanu hou no nā hana pili.

6. Hoʻohana pono ʻole i ka pahu.

Manaʻo Manaʻo: E kūʻai i kahi mea kanu DNA Isolation Kit i hoʻolaʻa ʻia i nā laʻana no ka unuhi ʻana a me ka hoʻomaʻemaʻe ʻana i ka DNA genomic mea kanu.

7. E hoʻopaʻa i ka WB me ka hoʻohui ʻole i kahinhydrous ethanol.

Manaʻo Manaʻo: E hōʻoia e hoʻohui i ka leo kūpono o ka ethanol piha i ka Buffer WB.

8. ʻAʻole i kulu pono ʻia ka eluent ma luna o ka membrane silica.

Manaʻo: E hoʻohui i ka eluent i hoʻomehana mua ʻia ma 65℃dropwise i ka waena o ka silica gel membrane, a waiho ia ma ka lumi wela no 5 minuke e hoonui i ka elution pono.

ʻO ka unuhi ʻana no ka loaʻa ʻana o ka DNA genomic haʻahaʻa

1. Ua mālama pono ʻole ʻia ka hāpana no ka lōʻihi loa, e hopena i ka hōʻino ʻana o ka DNA genomic.

Manaʻo Manaʻo: E mālama i nā laʻana kiko ma -20℃;e ho'āʻo e hoʻohana i nā laʻana kikoo hou i hōʻiliʻili ʻia no ka unuhi ʻana i ka DNA genomic.

2. Inā he liʻiliʻi loa ka nui o nā mea kikoo, e emi ana ka DNA genomic i unuhi ʻia.

Manaʻo Manaʻo: Ua waiwai kekahi mau mea kanu i ka wai, e like me nā mea kanu wai e like me ka algae, etc.

3. ʻAʻole i hoʻomaloʻo maikaʻi ʻia nā laʻana me ka naikokene wai a i ʻole waiho ʻia ma ka lumi wela no ka lōʻihi ma hope o ka wili ʻana.

Manaʻo Manaʻo: Pono e lawa ka wili naikokene wai, a ʻo ka paia o ke kelepona hāpana e wāwahi ʻia e like me ka hiki;ma hope koke o ka wili ʻana, pono e hoʻoneʻe ʻia ka pauka hāpana i 65℃preheated Buffer PL1 no ka hana aʻe.

4. ʻAʻole hoʻohana i ka pahu kūpono.

Manaʻo Manaʻo: E hoʻohana i ka Plant DNA Isolation Kit no ka unuhi a hoʻomaʻemaʻe i ka DNA genomic mea kanu.

5. ʻO ka mālama pono ʻole ʻana o Foregene Protease ka hopena i ka hoʻemi ʻana a i ʻole ka hana ʻole.

Manaʻo: E hōʻoia i nā kūlana mālama o Foregene Protease a i ʻole e hoʻololi iā ia me kahi Foregene Protease hou no ka hydrolysis enzymatic.

6. Pilikia maikaʻi

Manaʻo: E ʻoluʻolu e hoʻohana i ka Buffer EB no ka elution;inā hoʻohana ʻo ddH₂ʻO a i ʻole nā ​​mea eluent ʻē aʻe, e hōʻoia i ka pH o ka eluent ma waena o 7.0-8.5.

7. ʻAʻole kulu pololei ka eluent

Manaʻo Manaʻo: E ʻoluʻolu e hoʻohui i ka hāʻule elution i waenakonu o ka membrane silica a waiho i ka lumi wela no 5 mau minuke e hoʻonui ai i ka pono elution.

8. He liʻiliʻi loa ka leo eluent

Manaʻo: E ʻoluʻolu e hoʻohana i ka eluent no ka elution DNA genomic e like me nā kuhikuhi, ʻaʻole liʻiliʻi ma lalo o 100μl.

Ua unuhi ʻia ka DNA genomic me ka maʻemaʻe haʻahaʻa

ʻO ka haʻahaʻa haʻahaʻa o ka genomic DNA e alakaʻi i ka hāʻule ʻole a i ʻole ka hopena maikaʻi ʻole o nā hoʻokolohua o lalo, e like me: ʻaʻole hiki ke ʻoki ʻia ka enzyme, a ʻaʻole hiki ke loaʻa ka ʻāpana gene i ka PCR.

1. ʻO ka hoʻohaumia ʻana o nā protein ʻokoʻa, ka hoʻohaumia RNA.

Nānā: ʻAʻole i hoʻohana ʻia ʻo Buffer PW e holoi i ke kolamu;ʻAʻole i hoʻohana ʻia ʻo Buffer PW e holoi i ke kolamu ma ka wikiwiki centrifugation pololei.

Manaʻo: e hoʻāʻo e hōʻoia i ka ʻole o ka ua i loko o ka supernatant ke hele ka supernatant i ke kolamu;e holoi i ke kolamu hoʻomaʻemaʻe me Buffer PW e like me nā kuhikuhi, a ʻaʻole hiki ke haʻalele ʻia kēia hana.

2. Impurity ion pollution.

Nānā: Ua kāpae ʻia ke kolamu holoi WB Buffer a i ʻole holoi ʻia hoʻokahi wale nō, e hopena i ke koena ionic contamination.

Manaʻo Manaʻo: E holoi ʻelua me ka Buffer WB e like me nā ʻōlelo aʻoaʻo e wehe i nā koena ion e like me ka hiki.

3. ʻO ka haumia RNase.

Nānā: Hoʻohui ʻia ʻo Exogenous RNase i ka Buffer;ʻO ka hana holoi hewa ʻole ma Buffer PW e hopena i ke koena RNase a pili i nā hana hoʻokolohua RNA i lalo, e like me ka transcription in vitro.

Manaʻo: Foregene series nucleic acid extraction kits hiki ke wehe i ka RNA me ka loaʻa ʻole o RNase hou, a ʻaʻole pono nā reagents a pau i ka Plant DNA Isolation Kit i ka RNase;e holoi i ke kolamu hoʻomaʻemaʻe me Buffer PW e like me nā kuhikuhi, a ʻaʻole hiki ke haʻalele ʻia kēia hana.

4. Nā koena ethanol.

Nānā: Ma hope o ka holoi ʻana i ke kolamu hoʻomaʻemaʻe me Buffer WB, ʻaʻole i hana ʻia ka centrifugation paipu ʻole.

Manaʻo: E hahai i nā ʻōlelo aʻoaʻo no ka centrifugation paipu nele.

Papa kuhikuhi:

Ka Papa Hana Hoʻokaʻawale DNA Mea kanu