Igitabo cyo gusesengura ibibazo

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.

Umusaruro muke cyangwa nta ADN

Mubisanzwe hariho ibintu byinshi bigira ingaruka kumusaruro wa ADN genomic, harimo inkomoko yicyitegererezo, imyaka yicyitegererezo, imiterere yububiko bwikitegererezo, nigikorwa.

ADN ya genomic ntishobora kuboneka mugihe cyo kuyikuramo

1. Ingero za tissue zabitswe nabi cyangwa zibitswe igihe kirekire, bikaviramo kwangirika kwa ADN genomic.

Icyifuzo: Bika ingero za tissue muri azote yuzuye cyangwa -20°C;gerageza gukoresha ingero zegeranijwe zo gukuramo ADN genomic.

2. Umubare muto wicyitegererezo urashobora gutuma ADN ikomokaho idakurwa.

Icyifuzo: Kuburugero rwimitsi yabitswe igihe kirekire cyangwa ifite ADN yangirika cyane, umubare wintangarugero urashobora kwiyongera muburyo bukwiye kugirango ukuremo ADN nyinshi.Ingano yicyitegererezo irashobora kugenwa ukurikije ADN ikeneye, ariko icyitegererezo gishya ntigishobora kurenga 100mg, kandi icyitegererezo cyumye ntigishobora kurenga 30mg.

3. Icyitegererezo ntabwo kiri hamwe na azote yuzuye cyangwa ngo ishyirwe igihe kinini nyuma ya azote yuzuye.

Icyifuzo: Mugihe cyo gukuramo ADN, icyitegererezo kigomba kuba cyuzuye hamwe na azote yuzuye kugirango isenye urukuta rw'akagari;nyuma yo gusya, nyamuneka wohereze ifu yicyitegererezo kuri PL1 yashyutswe kuri 65°C vuba bishoboka (ifu yubutaka imaze gushonga, ADN genomic izatangira kwangirika vuba).

4. Ububiko budakwiye bwa Protegene Protease bivamo ibikorwa bigabanutse cyangwa bidakora.

Icyifuzo: Emeza uburyo bwo kubika Protease ya Foregene cyangwa uyisimbuze Protease nshya ya Foregene ya hydrolysis ya enzymatique.

5. Igikoresho kibitswe nabi cyangwa kibitswe igihe kirekire, gitera ibice bimwe mubikoresho kunanirwa.

Icyifuzo: Kugura ibikoresho bishya bya genomic ADN yo gukuramo ibikorwa bijyanye.

6. Gukoresha nabi ibikoresho.

Icyifuzo: Kugura Igiti cya ADN Yigunga Igikoresho cyahariwe ingero zo gukuramo no kweza ADN genomic.

7. Buffer WB utongeyeho anhydrous etanol.

Icyifuzo: Witondere kongeramo ingano yukuri ya Ethanol yuzuye kuri Buffer WB.

8. Umuhanga ntabwo yatonywe kuri silika membrane neza.

Igitekerezo: Ongeraho eluent yabanje gushyuha kuri 65℃kumanuka ugana hagati ya silika gel membrane, hanyuma ukayirekera mubushyuhe bwicyumba muminota 5 kugirango wongere imikorere ya elution.

Gukuramo kugirango ubone ADN itanga umusaruro muke

1. Icyitegererezo kibitswe nabi cyangwa kibitswe igihe kirekire, bikaviramo kwangirika kwa ADN genomic.

Icyifuzo: Bika ingero za tissue kuri -20℃;gerageza gukoresha ingero za tissue nshya zegeranijwe kugirango zivemo ADN.

2. Niba ingano yintangarugero ari nto cyane, ADN yakuweho izaba mike.

Icyifuzo: Bimwe mubitegererezo byibimera bikungahaye kumazi, nkibimera byo mumazi nka algae, nibindi, dosiye irashobora kwiyongera muburyo bukwiye cyangwa amazi ashobora kubura umwuma mbere gato yo gukora.

3. Ingero ntizari zifite neza na azote yuzuye cyangwa ngo zisigare ubushyuhe bwicyumba igihe kinini nyuma yo gusya.

Igitekerezo: Gusya kwa azote bigomba kuba bihagije, kandi urukuta rw'icyitegererezo rugomba kumeneka bishoboka;ako kanya nyuma yo gusya, ifu yicyitegererezo igomba kwimurwa kuri 65℃yashyutswe Buffer PL1 intambwe ikurikira.

4. Kudakoresha ibikoresho byiza.

Icyifuzo: Koresha ibikoresho byabigenewe byabigenewe ADN yo kwigunga kugirango ukure kandi usukure ADN yibimera.

5. Ububiko budakwiye bwa Foregene Protease bivamo ibikorwa bigabanutse cyangwa bidakora.

Icyifuzo: Emeza uburyo bwo kubika Protease ya Foregene cyangwa uyisimbuze Protease nshya ya Foregene ya hydrolysis ya enzymatique.

6. Ikibazo gikomeye

Icyifuzo: Nyamuneka koresha Buffer EB kugirango ukureho;niba ukoresha ddH₂O cyangwa ibindi bisobanuro, menya neza ko pH ya eluent iri hagati ya 7.0-8.5.

7. Umuhanga ntabwo yatonywe neza

Igitekerezo: Nyamuneka ongeraho igitonyanga cya elution hagati ya silika membrane hanyuma ubirekere mubushyuhe bwicyumba muminota 5 kugirango wongere imikorere ya elution.

8. Ingano ya eluent ni nto cyane

Igitekerezo: Nyamuneka koresha eluent mugukuraho ADN genomic ukurikije amabwiriza, byibuze bitarenze 100μl.

Yakuweho ADN genomic ifite isuku nke

Ubuziranenge buke bwa ADN genomic bizatera kunanirwa cyangwa ingaruka mbi zubushakashatsi bwo hasi, nka: enzyme ntishobora kugabanywa, kandi agace ka gene ntigashobora kuboneka na PCR.

1. Kwanduza poroteyine zitandukanye, kwanduza RNA.

Isesengura: Buffer PW ntabwo yakoreshejwe mu koza inkingi;Buffer PW ntabwo yakoreshejwe mu koza inkingi kumuvuduko ukwiye wa centrifugation.

Igitekerezo: gerageza urebe ko nta mvura igwa muri ndengakamere iyo ndengakamere inyuze mu nkingi;menya neza koza inkingi yo kweza hamwe na Buffer PW ukurikije amabwiriza, kandi iyi ntambwe ntishobora kuvaho.

2. Guhumanya ion.

Isesengura: Buffer WB yo gukaraba inkingi yasibwe cyangwa yogejwe rimwe gusa, bivamo kwanduza ionic.

Icyifuzo: Witondere gukaraba kabiri hamwe na Buffer WB ukurikije amabwiriza yo gukuraho ion zisigaye bishoboka.

3. Kwanduza RNase.

Isesengura: Exogenous RNase yongewe kuri Buffer;ibikorwa byo gukaraba nabi muri Buffer PW bizavamo RNase isigaye kandi bigire ingaruka kumikorere ya RNA yo hepfo, nko muri vitro transcription.

Igitekerezo: Ibikoresho byo gukuramo nucleic aside ya Foregene birashobora gukuramo RNA nta RNase yongeyeho, kandi reagent zose ziri muri Plant DNA Isolation Kit ntizikeneye RNase;menya neza koza inkingi yo kweza hamwe na Buffer PW ukurikije amabwiriza, kandi iyi ntambwe ntishobora kuvaho.

4. Ibisigazwa bya Ethanol.

Isesengura: Nyuma yo koza inkingi yo kweza hamwe na Buffer WB, nta centrifugation yubusa yakozwe.

Icyifuzo: Kurikiza amabwiriza yuburyo bukwiye bwa centrifugation.

Igitabo gikubiyemo amabwiriza:

Tera ADN Igitabo cyo Kwifashisha Igitabo