Ukuhlaziywa okulandelayo kwezinkinga ezingenzeka kuBuccalswab/I-FTA card DNA extraction is helpful to your experiment. 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.

Ikholomu yokuhlanza ivalekile

Kule kit, ekusebenzeni kokukhipha i-DNA ye-genomic, ikholomu yokuhlanza ikhangiswa ngokuqondile kungxube yesampula ye-enzymatic lysis ngaphandle kwesinyathelo se-centrifugation, futhi ikholomu yokuhlanza ingase ivinjwe ngenxa ye-enzymization engaphelele kanye ne-viscosity ephezulu yesampula.

Izimbangela ezilandelayo zingaba kanje:

1. Ukugaya okungaphelele kwe-enzymatic kwamasampula ezicubu.

Isincomo: Isikhathi sokucubungula isampula se-Foregene Protease singanwetshwa ngokufanelekile noma i-supernatant ingathathwa ngemva kokumiswa maphakathi ngo-12,000 rpm (~13,400× g) imizuzu emi-5.

2. Ukusetshenziswa ngokweqile kwamasampula ezicubu noma izicubu ezinkulu.

Isincomo: Kungcono ukuthi ungadluli ku-1Buccal swab kusampula;uma isampula inkulu kakhulu, khulisa umthamo we-Buffer ST1, Foregene Protease, buffer ST2 ngokufanele.

3. I-viscosity yesampula iphezulu kakhulu.

Isincomo: Amasampuli angahlanjululwa ngokufanelekile ngo-10 mM we-Tris-HCl ngaphambi kokukhipha i-genomic DNA.

4. Izingcezwana zekhadi legazi zimunyiwe.

Okutuswayo: Isikhathi esidlulayo se-centrifugation sesinyathelo sesi-6 sendawo yegazi (Ikhadi le-FTA) lokukhishwa kwe-genomic singanwetshwa ngokufanelekile.

Isivuno esiphansi noma ayikho i-DNA

Kuvame ukuba nezinto ezihlukahlukene ezithinta isivuno se-genomic DNA, okuhlanganisa imvelaphi yesampula, izimo zokugcinwa kwesampula, ukulungiswa kwesampula, ukukhohlisa, njll.

I-Genomic DNA ayikwazi ukutholwa ngesikhathi sokukhishwa

Izimbangela ezingenzeka zimi kanje:

1. Ukulondolozwa okungafanele kwamasampuli noma ukugcinwa isikhathi eside kuholela ekonakaleni kwe-DNA ye-genomic.

Okutuswayo: Ama-swabs omlomo kufanele athathwe njengesampula entsha, futhi akufanelekile ukusebenzisa ama-swabs alondoloziwe ukuze kwenziwe imisebenzi yokukhipha i-DNA ye-genomic;amasampula amachashaza egazi kufanele aqinisekise ukuthi ikhwalithi ifanelekile futhi isikhathi sokugcina akufanele sibe side kakhulu.

2. Ukusetshenziswa kwezicubu ezincane kakhulu kungase kuphumele ekukhishweni kwe-DNA ye-genomic ehambisanayo.

Isincomo: Landela i-bhekal imiyalo yesampula ye-swab kumhlahlandlela wokusebenza, futhi usule izikhathi eziningi ngangokunokwenzeka ukuze amaseli anele akwazi ukunamathiselwa ku-oral swab ukuze kukhishwe i-genomic DNA;ukuze kukhishwe isampula yendawo yegazi, indawo yokusika indawo yegazi ingakhuliswa ngokufanelekile.

3. I-Foregene Protease ayilondolozwa ngendlela engafanele, okuholela ekwehleni komsebenzi wayo noma ukungasebenzi.

Isincomo: Qinisekisa izimo zokugcina zei-Fi-oregene Protease noma esikhundleni sayo ufake i-Foregene Protease entsha yokusabela kwe-enzymatic.

4. Ukulondolozwa okungalungile kwekhithi noma isikhathi sokugcina side kakhulu, okuholela ekuhlulekeni kwezinye izingxenye zekhithi.

Okunconyiwe: Thenga okushaBuccal swab DNAukuzihlukanisa ikhithi yezinqubo ezihlobene.

5. I-Buffer WB ayifaki i-ethanol ephelele.

Isincomo: Qinisekisa ukuthi isigcinalwazi i-WB sengeza ivolumu elungile ye-ethanol ephelele.

6. I-eluent ayifakwanga kahle kwifilimu ye-silicone.

Isincomo: Engeza 65°CI-eluent efudunyeziwe yehla iye maphakathi nolwelwesi lwe-silicone futhi ishiye ekamelweni lokushisa imizuzu emi-5 ukuze kwandiswe ukusebenza kahle kwe-elution.

Li-ow-yield genomic DNA eyedwa

Izimbangela ezilandelayo zingaba kanje:

1. Ukulondolozwa okungafanele kwamasampuli noma ukugcinwa isikhathi eside kuholela ekonakaleni kwe-DNA ye-genomic.

Okutuswayo: Ama-swabs omlomo kungcono athathwe amasampula amasha, futhi ama-swabs alondoloziwe akufanele asetshenziselwe ukukhipha i-DNA ye-genomic.

2. Uma inani lesampula lethishu lilincane kakhulu, okuqukethwe kwe-DNA ekhishiwe kuzoba kuncane.

Okutuswayo: Landela imiyalelo yesampula ye-oral swab kumhlahlandlela wokusebenza, wesula izikhathi eziningi ngangokunokwenzeka ukuze amaseli anele anamathiselwe ku-oral swab ukuze kukhishwe i-genomic DNA.

3. I-Foregene Protease ayilondolozwa ngendlela engafanele, okuholela ekwehleni komsebenzi wayo noma ukungasebenzi.

Isincomo: Qinisekisa izimo zokugcina zethe Fi-oregene Protease noma uyishintshe ufake entsha I-Foregene Protease yokusabela kwe-enzymatic.

4. Izinkinga ze-Eluent.

Isincomo: Sebenzisa i-Buffer EB ukuze uthole i-lution;uma usebenzisa i-ddH₂O noma ezinye izibonelo, qinisekisa ukuthi i-pH ye-eliate iphakathi kuka-7.0-8.5.

5. I-eluate ayifakwanga ngendlela efanele.

Okutuswayo: Engeza amaconsi alula phakathi nolwelwesi lwe-silicone bese ushiya ekamelweni lokushisa imizuzu emi-5 ukuze ukwandise ukusebenza kahle kwe-elution.

6. Uketshezi lwe-elution lunqwabelana luncane kakhulu.

Okunconyiwe: Sebenzisa i-eluent ye-genomic DNA elution njengoba kudingwa emiyalweni, okungenanihhayi ngaphansi ezingaphezu kuka-15μl.

Lubumsulwa of i-genomic DNAeyedwa

Ukuhlanzeka okuphansi kwe-DNA ye-genomic kungaholela ekuhlulekeni noma emiphumeleni engagculisi yokuhlola okuya phansi, okufana nalokhu: ama-enzyme awakwazi ukusikwa avuleke, i-PCR ayikwazi ukuthola ucezu lofuzo oluthakaselwayo, njll.

Izimbangela ezingenzeka zimi kanje:

1. Ukungcola kwe-Heteroprotein, ukungcola kwe-RNA.

Ukuhlaziya: Ikholomu yokuhlanza ayizange igezwe kusetshenziswa i-Buffer PW;ikholomu yokuhlanza i-Buffer PW ayizange iwashwe kusetshenziswa isivinini esilungile se-centrifugal.

Okutuswayo: Qinisekisa ukuthi akukho mvula kumandla amakhulu ngaphambi kokwengeza i-ethanol;qiniseka ukuthi ugeza ikholomu yokuhlanza ngokusho kwemiyalelo, futhi lesi sinyathelo asikwazi ukushiywa.

2. Ukungcoliswa kwe-ion engcolile.

Ukuhlaziywa: Ikholomu yokuhlanza i-Buffer WB iye yashiywa noma yawashwa kanye kuphela, okuholele ekungcoleni okusele kwe-ionic.

Okutuswayo: Qiniseka ukuthi uyayigeza i-Buffer WB izikhathi ezi-2 njengoba uyalelwe ukuze ususe ama-ion ayizinsalela ngangokunokwenzeka.

3. Ukungcola kwe-enzyme ye-RNA.

Ukuhlaziya: Ama-RNa angaphandle angeziwe ku-buffer;Ukusebenza kokugeza kwe-Buffer PW bekungalungile, okubangele izinsalela ze-RNase, kukhinyabeze imisebenzi yokuhlola ye-RNA esezansi nomfula, njengokulotshwa kwe-in vitro.

Isincomo: Foregene series nucleic acidisolaamakhithi e-tion angasusa i-RNA ngaphandle kokwengezwa okwengeziwe kwe-RNase,kanjalo i-buccal Swab/FTA Card DNA Isolation Kitisidingo't engeza i-RNase;qiniseka ukuthi ulandela imiyalelo yekholomu yokuhlanza i-Buffer PW, futhi lesi sinyathelo asinakushiywa.

4. Izinsalela ze-Ethanol.

Ukuhlaziywa: I-Buffer WB ayizange yenze i-tube centrifugation engenalutho ngemva kokugeza ikholomu yokuhlanza.

Isincomo: Yenza umsebenzi olungile we-tube centrifugation ngokulandela imiyalelo.

5. Okunye ukungcola kokungcola.

Ukuhlaziya: Amasampuli alondoloziwe noma amasampuli akhethekile awaphathwa kusengaphambili.

Okutuswayo: Yilungise kahle isampula njengoba uyaliwe.