Olu hlalutyo lulandelayo lweengxaki ezinokwenzeka kwiBuccalswab/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.

Ikholamu yokucoca ivaliwe

Kule kit, kwi-genomic DNA extraction operation, ikholomu yokucoca i-adsorbed ngokuthe ngqo kumxube we-sampulu ye-enzymatic lysis ngaphandle kwesinyathelo se-centrifugation, kwaye ikholamu yokucoca inokuthi ivalwe ngenxa ye-enzymization engaphelelanga kunye ne-viscosity ephezulu yesampuli.

Ezi zinto zilandelayo zinokuba ngunobangela zezi zilandelayo:

1. I-enzymatic engaphelelanga yokugaya iisampuli zezicubu.

Isindululo: Ixesha lokucubungula isampulu ye-Foregene Protease inokwandiswa ngokufanelekileyo okanye i-supernatant ingathathwa emva kwe-centrifugation kwi-12,000 rpm (~13,400× g) imizuzu emi-5.

2. Ukusetyenziswa ngokugqithiseleyo kweesampuli zezicubu okanye izicubu ezinkulu.

Ingcebiso: Kungcono ukuba ungadluli ku-1Buccal swab kwisampuli;ukuba isampulu inkulu kakhulu, nyusa idosi yeBuffer ST1, Foregene Protease, buffer ST2 ngokufanelekileyo.

3. Isampula ye-viscosity iphezulu kakhulu.

Ingcebiso: Iisampulu zinokuxutywa ngokufanelekileyo nge-10 mM ye-Tris-HCl phambi kokutsalwa kwe-genomic DNA.

4. Iziqwenga zekhadi legazi ziye zamuncwa.

Ingcebiso: Ixesha elidlulayo le-centrifugation yenyathelo lesi-6 lendawo yegazi (iKhadi le-FTA) i-genomic extraction inokwandiswa ngokufanelekileyo.

Isivuno esiphantsi okanye akukho DNA

Ngokuqhelekileyo kukho izinto ezahlukeneyo ezichaphazela isivuno se-genomic DNA, kubandakanywa imvelaphi yesampuli, iimeko zokugcina isampuli, ukulungiswa kwesampuli, ukukhwabanisa, njl.

I-Genomic DNA ayinakufumaneka ngexesha lokukhutshwa

Izizathu ezinokuthi zibe zezi zilandelayo:

1. Ukugcinwa okungafanelekanga kweesampuli okanye ukugcinwa kwexesha elide kukhokelela ekuthotyweni kwe-genomic DNA.

Isincomo: I-swabs yomlomo kufuneka ikhethwe ngokutsha, kwaye akucebisi ukusebenzisa i-swabs egciniweyo kwimisebenzi ye-genomic DNA extraction;iisampuli zendawo yegazi kufuneka ziqinisekise ukuba umgangatho ufanelekile kwaye ixesha lokugcina akufanele libe lide kakhulu.

2. Ukusetyenziswa kwezicubu ezincinci kunokubangela ukuba kungabikho kukhutshwa kwe-DNA ye-genomic ehambelana nayo.

Ingcebiso: Landela ibuccal i-swab yesampula imiyalelo kwisikhokelo sokusebenza, kwaye usule amaxesha amaninzi kangangoko ukuze iiseli ezaneleyo zifakwe kwi-oral swab ye-genomic DNA extraction;Ukutsalwa kwesampulu yendawo yegazi, indawo yokusika indawo yegazi inokongezwa ngokufanelekileyo.

3. I-Foregene Protease ayigcinwanga ngokufanelekileyo, okukhokelela ekunciphiseni komsebenzi wayo okanye ukungasebenzi.

Ingcebiso: Qinisekisa iimeko zokugcina zeiFi-oregene Protease okanye endaweni yayo ufake i-Foregene Protease entsha ye-enzymatic reaction.

4. Ukugcinwa okungafanelekanga kwekiti okanye ixesha lokugcina ixesha elide kakhulu, okubangelwa ukungaphumeleli kwamanye amacandelo kwikiti.

Ingcebiso: Thenga entshaBuccal swab DNAukubekwa wedwa ikhithi yeenkqubo ezinxulumeneyo.

5. I-Buffer WB ayongezi i-ethanol ngokupheleleyo.

Ingcebiso: Qinisekisa ukuba i-buffer WB yongeza umthamo ochanekileyo we-ethanol epheleleyo.

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

Ingcebiso: Yongeza 65°CI-eluent efudunyeziweyo iwa phakathi kwe-membrane ye-silicone kwaye ishiye kwiqondo lobushushu le-5 min ukunyusa ukusebenza kakuhle kwe-elution.

Li-ow-yield genomic DNA kwadwa

Ezi zinto zilandelayo zinokuba ngunobangela zezi zilandelayo:

1. Ukugcinwa okungafanelekanga kweesampuli okanye ukugcinwa kwexesha elide kukhokelela ekuthotyweni kwe-genomic DNA.

Isincomo: Iiswabhu zomlomo zikhethwa ukuba zisanda kuthathwa iisampulu, kwaye i-swabs egciniweyo ayifanele isetyenziselwe ukutsalwa kwe-genomic DNA.

2. Ukuba inani lesampula yezicubu lincinci kakhulu, umxholo we-DNA okhutshweyo uya kuba ngaphantsi.

Isincomo: Landela imiyalelo yesampulu ye-oral swab kwisikhokelo sokusebenza, usule amaxesha amaninzi kangangoko ukuze iiseli ezaneleyo zifakwe kwi-oral swab ye-genomic DNA extraction.

3. I-Foregene Protease ayigcinwanga ngokufanelekileyo, okukhokelela ekunciphiseni komsebenzi wayo okanye ukungasebenzi.

Ingcebiso: Qinisekisa iimeko zokugcina zethe Foregene Protease okanye endaweni yayo ufake entsha I-Foregene Protease ye-enzymatic reaction.

4. Iingxaki ze-Eluent.

Ingcebiso: Sebenzisa i-Buffer EB yolution;ukuba usebenzisa i-ddH₂O okanye ezinye ii-eluents, ziqinisekisa ukuba i-pH ye-eluate iphakathi kwe-7.0-8.5.

5. I-eluate ayidityaniswanga ngokuchanekileyo.

Isincomo: Yongeza i-eluent drops kumbindi we-membrane ye-silicone kwaye ushiye kwindawo yobushushu bemizuzu emi-5 ukwandisa ukusebenza kakuhle kwe-elution.

6. Ulwelo lwe-elution luqokelelana kancinci kakhulu.

Ingcebiso: Sebenzisa i-eluent ye-genomic DNA elution njengoko kufunwa kwimiyalelo, ubuncincihayi ngaphantsi ngaphezu kwe-15μl.

Lubunyulu of i-DNA ye-genomickwadwa

Ukucoceka kwe-DNA ye-genomic ephantsi kunokukhokelela ekungaphumeleli okanye kwiziphumo ezingonelisiyo zezilingo ezisezantsi, ezifana: i-enzymes ayinakunqunyulwa ivuleke, i-PCR ayikwazi ukufumana i-gene fragment of interest, njl.

Izizathu ezinokuthi zibe zezi zilandelayo:

1. Ungcoliseko lweHeteroprotein, ungcoliseko lweRNA.

Uhlalutyo: Ikholamu yokucoca ayizange ihlanjwe ngokusebenzisa i-Buffer PW;i-Buffer PW yokuhlamba ikholamu ayizange ihlanjwe kusetyenziswa isantya esichanekileyo se-centrifugal.

Ingcebiso: Qinisekisa ukuba akukho mvula kwi-supernatant phambi kokuba wongeze i-ethanol;qiniseka ukuba uhlamba ikholamu yokucoca ngokwemiyalelo, kwaye le nyathelo ayinakushiywa.

2. Ungcoliseko lwe-ion.

Uhlalutyo: Ikholamu yokucoca i-Buffer WB iye yashiywa okanye yahlanjwa kube kanye kuphela, okubangele ukungcoliseka kwe-ionic okushiyekileyo.

Ingcebiso: Qiniseka ukuba uyayihlamba i-Buffer WB ka-2 njengoko uyalelwe ukususa iiyoni ezishiyekileyo kangangoko kunokwenzeka.

3. Ungcoliseko lwe-enzyme ye-RNA.

Uhlalutyo: Ii-RNa zangaphandle zongezwa kwi-buffer;Umsebenzi wokuhlamba we-Buffer PW wawungalunganga, ubangele iintsalela ze-RNase, ezichaphazela imisebenzi yovavanyo ye-RNA esezantsi, efana noshicilelo lwe-in vitro.

Isincomo: Foregene series nucleic acidisolaIikiti ze-tion zinokususa i-RNA ngaphandle kokongeza i-RNase,ngolo hlobo buccal Swab / FTA Card DNA Isolation Kitisidingo't yongeza i-RNase;qiniseka ukuba ulandela imiyalelo yeKholam yokucoca i-Buffer PW, kwaye eli nyathelo alinako ukushiyeka.

4. Intsalela ye-Ethanol.

Uhlalutyo: I-Buffer WB ayizange yenze i-tube centrifugation engenanto emva kokuhlamba ikholamu yokucoca.

Isincomo: Yenza umsebenzi ochanekileyo wetyhubhu engenanto ye-centrifugation ngokwemiyalelo.

5. Olunye ungcoliseko lokungcola.

Uhlalutyo: Iisampulu ezigciniweyo okanye iisampulu ezikhethekileyo azilungiswa.

Ingcebiso: Yilungise kakuhle isampuli njengoko uyalelwe.