Plant DNA Isolation Kit Genomic Plant DNA Purification Kits Reagents Protocol
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Le khithi isebenzisa ikholomu ye-DNA kuphela engabopha ngokuqondile i-DNA, i-Foregene protease kanye nesistimu ye-buffer eyingqayizivele, eyenza kube lula kakhulu ukuhlanzwa kwe-DNA genomic yezitshalo.Ikhwalithi ephezulu ye-genomic DNA ingatholakala phakathi nemizuzu engama-30, egwema ukucekelwa phansi kwe-genomic DNA.
Ulwelwesi lwejeli ye-silica ye-DNA kuphela esetshenziswa kukholamu yokuphotha iyinto entsha eyingqayizivele ye-Foregene, engabopha ngempumelelo futhi ngokuqondile ku-DNA, futhi yandise ukukhishwa kwe-RNA, amaprotheni angcolile, ama-ion, ama-polysaccharides, ama-polyphenols nezinye izinhlanganisela eziphilayo.
Izingxenye zomkhiqizo
| I-Buffer PL1, Buffer PL2 |
| I-Buffer PW, Buffer WB, Buffer EB |
| I-Foregene Protease |
| Ikholomu Ye-DNA Kuphela |
| Iziyalezo |
Izici&izinzuzo
■ Akukho ukungcoliswa kwe-RNase: Ikholomu Ye-DNA Kuphela ehlinzekwa ikhithi yenza kube nokwenzeka ukususa i-RNA ku-DNA ye-genomic ngaphandle kwe-RNase eyengeziwe phakathi nokuhlolwa, ukuvimbela ilabhorethri ukuthi ingangcoliswa i-RNase yangaphandle.
■ Isivinini esisheshayo: I-Foregene Protease inomsebenzi ophezulu kunama-protease afanayo futhi igaya amasampula ezicubu ngokushesha.
■ Okulula: ukusebenza kwe-genomic DNA extraction kungaqedwa ngemizuzu engama-30.
■ Ilula: I-centrifugation yenziwa ekamelweni lokushisa, akukho 4℃ lokushisa eliphansi centrifugation noma ethanol precipitation of DNA.
■ Ukuphepha: asikho i-reagent ephilayo edingekayo.
■ Ikhwalithi ephezulu: I-DNA ehlanzekile ye-genomic inezingcezu ezinkulu, ayikho i-RNA, ayikho i-RNase, kanye nokuqukethwe kwe-ion ephansi kakhulu, kungahlangabezana nezidingo zokuhlola okuhlukahlukene.
Isicelo sekhithi
Ifanele ukukhishwa nokuhlanzwa kwe-genomic DNA ezicutshini zezitshalo ezintsha noma eziqandisiwe.
Ukugeleza komsebenzi
Umdwebo
Isitoreji kanye Nempilo yeshelufu
Ikhithi ingagcinwa izinyanga eziyi-12 ekamelweni lokushisa (15-25 ℃) kanye no-2–8℃ isikhathi eside.
Isixazululo se-Foregene Protease Plus sinefomula eyingqayizivele, esebenzayo uma igcinwe ekamelweni lokushisa isikhathi eside (izinyanga ezi-3);umsebenzi wayo nokuzinza kuzoba ngcono uma igcinwe4℃, ngakho-ke kunconywa ukuthi uyigcine ku-4℃, khumbula ukuthi ungayigcini ku -20℃.
Umhlahlandlela Wokuhlaziya Inkinga
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.
Isivuno esiphansi noma ayikho i-DNA
Ngokuvamile kuba nezinto eziningi ezithinta umkhiqizo we-DNA ye-genomic, okuhlanganisa umthombo wesampula, iminyaka yesampula, izimo zokugcinwa kwesampula, kanye nokusebenza.
I-Genomic DNA ayitholakalanga ngesikhathi sokukhishwa
1. Amasampula ezicubu agcinwa ngendlela engafanele noma agcinwe isikhathi eside kakhulu, okuholela ekulimaleni kwe-DNA ye-genomic.
Okutuswayo: Gcina amasampula ezicubu ku-nitrogen ewuketshezi noma -20°C;zama ukusebenzisa amasampula asanda kuqoqwa ukuze kukhishwe i-genomic DNA.
2. Inani elincane kakhulu lesampula lingabangela ukuthi i-DNA ye-genomic ehambisanayo ingakhishwa.
Isiphakamiso: Kumasampula ezicubu agcinwe isikhathi eside noma anokonakala okukhulu kwe-genomic DNA, inani lamasampula ezicubu linganyuswa ngokufanelekile ukuze kukhishwe i-DNA ye-genomic.Inani lesampula linganqunywa ngokwezidingo ze-DNA, kodwa isampula esisha akufanele sidlule u-100mg, futhi isampula elomile akufanele lidlule u-30mg.
3. Isampula ayigaywa nge-nitrogen ewuketshezi noma ibekwe isikhathi eside kakhulu ngemva kwe-nitrogen ewuketshezi.
Isiphakamiso: Ngesikhathi sokukhishwa kwe-DNA, isampula idinga ukugaywa ngokugcwele nge-nitrogen ewuketshezi ukuze iphule udonga lweseli;ngemva kokugaya, sicela udlulisele impushana yesampula ku-PL1 eshisiwe ngo-65°C ngokushesha ngangokunokwenzeka (uma i-powder yomhlabathi incibilikile, i-DNA ye-genomic izoqala ukuwohloka ngokushesha) .
4. Ukugcinwa okungalungile kwe-Foregene Protease kubangela umsebenzi oncishisiwe noma ongasebenzi.
Okutuswayo: Qinisekisa izimo zokugcina ze-Foregene Protease noma ufake esikhundleni sayo i-Foregene Protease entsha ye-enzymatic hydrolysis.
5. Ikhithi igcinwa ngendlela engafanele noma igcinwe isikhathi eside kakhulu, okwenza ezinye izingxenye zekhithi zihluleke.
Okunconyiwe: Thenga ikhithi entsha yokukhipha i-genomic DNA yesitshalo ukuze uthole imisebenzi ehlobene.
6. Ukusetshenziswa ngendlela engafanele kwekhithi.
Isiphakamiso: Thenga I-Plant DNA Isolation Kit enikezelwe kumasampuli ukuze kukhishwe futhi kuhlanzwe i-plant genomic DNA.
7. Isikhafu WB ngaphandle kokwengeza aengenamanzi i-ethanol.
Isincomo: Qiniseka ukuthi ungeza ivolumu elungile ye-ethanol ephelele ku-Buffer WB.
8. I-eluent ayizange iconsele kulwelwesi lwesilika ngendlela efanele.
Ukusikisela: Engeza i-eluent eshisiwe ngaphambili ku-65℃i-dropwise iye maphakathi nolwelwesi lwejeli ye-silica, futhi uyishiye ekamelweni lokushisa imizuzu emi-5 ukuze ukwandise ukusebenza kahle kwe-elution.
Isizinda sokuthola i-DNA ye-genomic enesivuno esincane
1. Isampula ligcinwe ngendlela engafanele noma ligcinwe isikhathi eside kakhulu, okuholela ekulimaleni kwe-DNA ye-genomic.
Okutuswayo: Gcina amasampula ezicubu ku -20℃;zama ukusebenzisa amasampula ezicubu ezisanda kuqoqwa ukuze kukhishwe i-genomic DNA.
2. Uma inani lamasampula ezicubu lilincane kakhulu, i-DNA ye-genomic ekhishiwe izoba ngaphansi.
Ukusikisela: Amanye amasampula ezitshalo anothile ngamanzi, njengezitshalo zasemanzini ezifana nolwelwe, njll., umthamo unganyuswa ngokufanelekile noma amanzi angaphelelwa amanzi kancane ngaphambi kokuhlinzwa.
3. Amasampula awazange agaywe kahle nge-nitrogen ewuketshezi noma ashiywe ekamelweni lokushisa isikhathi eside kakhulu ngemva kokugaya.
Ukusikisela: Ukugaya i-nitrogen ewuketshezi kufanele kube ngokwanele, futhi udonga lweseli lwesampula kufanele luphuke ngangokunokwenzeka;ngokushesha ngemva kokugaya, isampula powder kufanele idluliselwe ku-65℃ishisa ngaphambili i-Buffer PL1 ngesinyathelo esilandelayo.
4. Ukungasebenzisi ikhithi efanele.
Isincomo: Sebenzisa i-Plant DNA Isolation Kit ezinikele ukuze ukhiphe futhi uhlanze i-genomic DNA yezitshalo.
5. Ukugcinwa okungalungile kwe-Foregene Protease kubangela umsebenzi oncishisiwe noma ongasebenzi.
Okutuswayo: Qinisekisa izimo zokugcina ze-Foregene Protease noma ufake esikhundleni sayo i-Foregene Protease entsha ye-enzymatic hydrolysis.
6. Inkinga ye-Eluent
Isincomo: Sicela usebenzise i-Buffer EB ukuthola ulwazi;uma usebenzisa i-ddH2O noma ezinye izibonelo, qiniseka ukuthi i-pH ye-eluent iphakathi kuka-7.0-8.5.
7. I-eluent ayiconsi kahle
Ukusikisela: Sicela wengeze ukwehla kwe-elution phakathi nolwelwesi lwe-silica futhi ukushiye endaweni efudumele imizuzu emi-5 ukuze ukwandise ukusebenza kahle kwe-elution.
8. Ivolumu yomsindo incane kakhulu
Ukusikisela: Sicela usebenzise i-eluent ukuze uthole i-genomic DNA elution ngokwemiyalelo, okungenani hhayi ngaphansi kwe-100.μl.
I-DNA ye-genomic ekhishwe ngobumsulwa obuphansi
Ukuhlanzeka okuphansi kwe-DNA ye-genomic kuzoholela ekuhlulekeni noma emphumeleni omubi wokuhlola okwehla nomfula, njengokuthi: i-enzyme ayikwazi ukunqunywa, futhi ucezu lofuzo oluhlosiwe alukwazi ukutholwa yi-PCR.
1. Ukungcola kwamaprotheni Okuxubile, ukungcola kwe-RNA.
Ukuhlaziywa: I-Buffer PW ayizange isetshenziselwe ukugeza ikholomu;I-Buffer PW ayizange isetshenziselwe ukugeza ikholomu ngesivinini esilungile se-centrifugation.
Ukusikisela: zama ukuqinisekisa ukuthi akukho mvula kumandla angaphezu kwavamile lapho amandla angaphezu kwavamile edlula kukholamu;qiniseka ukuthi ugeza ikholomu yokuhlanza nge-Buffer PW ngokwemiyalelo, futhi lesi sinyathelo asinakushiywa.
2. Ukungcoliswa kwe-ion engcolile.
Ukuhlaziywa: Ikholomu ye-Buffer WB yokugeza iye yashiywa noma yawashwa kanye kuphela, okuholele ekungcoleni okusalile kwe-ionic.
Okutuswayo: Qiniseka ukuthi ugeza kabili nge-Buffer WB ngokwemiyalelo ukuze ususe ama-ion ayinsalela ngangokunokwenzeka.
3. Ukungcola kwe-RNase.
Ukuhlaziywa: I-RNase Engaphandle yengezwe ku-Buffer;ukusebenza okungalungile kokuwasha ku-Buffer PW kuzoholela ku-RNase eyinsalela futhi kuthinte imisebenzi yokuhlola ye-RNA ezansi nomfula, njengokulotshwa kwe-in vitro.
Isiphakamiso: Ikhithi yokukhipha i-nucleic acid yochungechunge lwe-Foregene ingasusa i-RNA ngaphandle kwe-RNase eyengeziwe, futhi wonke ama-reagents ku-Plant DNA Isolation Kit awayidingi i-RNase;qiniseka ukuthi ugeza ikholomu yokuhlanza nge-Buffer PW ngokwemiyalelo, futhi lesi sinyathelo asinakushiywa.
4. Izinsalela ze-ethanol.
Ukuhlaziya: Ngemva kokugeza ikholomu yokuhlanza nge-Buffer WB, awekho amashubhu angenalutho okufakwa phakathi kwe-centrifugation okwenziwa.
Okunconyiwe: Landela imiyalelo ye-tube centrifugation efanele engenalutho.
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