Buccal Swab/FTA Card DNA Isolation Kit Genomic DNA Extraction okanye Purifiction Kit from Buccal Swabs
Inkcazelo yeKit:
Ngokukhawuleza uhlambulule i-DNA ye-genomic ephezulu kwi-buccal swab / iisampulu zeKhadi le-FTA.
◮Akukho susulelo lwe-RNase:IKholamu ye-DNA-Kuphela enikezelwe yikiti yenza kube nokwenzeka ukususa i-RNA kwi-DNA ye-genomic ngaphandle kokongeza i-RNase ngexesha lovavanyo, ukuthintela ilabhorethri ukuba ingangcoliswa yi-RNase yangaphandle.
◮Isantya esiphezulu:I-Foregene Protease inomsebenzi ophezulu kuneeproteases ezifanayo, kwaye yetyise iisampulu zethishu ngokukhawuleza;ukusebenza kulula, kwaye i-genomic DNA extraction operation ingagqitywa kwimizuzu engama-20-80.
◮Iluncedo:I-centrifugation yenziwa kwiqondo lokushisa, kwaye akukho mfuneko ye-4 ° C ephantsi kwe-centrifugation okanye i-ethanol precipitation ye-DNA.
◮Ukhuseleko:Akukho kukhutshwa kwe-organic reagent efunekayo.
◮Umgangatho ophezulu:I-DNA ekhutshiweyo ye-genomic inamaqhekeza amakhulu, akukho RNA, akukho RNase, kunye nomxholo we-ion ophantsi kakhulu, onokuhlangabezana neemfuno zeemvavanyo ezahlukeneyo.
◮Inkqubo yeMicro-elution:Inokunyusa uxinaniso lwe-DNA ye-genomic, elungele ukufunyanwa okanye ukuvavanywa komlambo osezantsi.
Inkcazo
Le kit ibonelela ngendlela esebenzayo kwaye ekhawulezayo yokufumana i-DNA ye-genomic ephezulu yoxinaniso kwi-buccal swabs kunye neKhadi le-FTA (amabala egazi).Ukusebenzisa inkampani yethu'I-s unique DNA-kuphela kwe-silica membrane spin ikholamu kunye nefomula, idibene ne-Foregene Protease, i-high-concentration, i-high-quality genomic DNA inokukhutshwa kwimizuzu ye-80.Ikholamu encinci yokucoca eyenzelwe ngokukodwa ibophelela i-DNA ye-genomic, kwaye i-DNA inokukhutshwa ngexabiso elincinci (15μl) inkqubo elution ukunyusa uxinaniso lwe-DNA ye-genomic efunyenweyo, elungele ukubhaqwa komlambo okanye umfuniselo.Ikiti inokwenza isampuli enye okanye ngaphezulu ngexesha, kwaye inkqubo yokucoca ayifuni ukutsalwa kwezinto eziphilayo ezifana ne-phenol, i-chloroform, kunye ne-isopropanol echitha ixesha okanye i-ethanol precipitation, kwaye umsebenzi ulula kwaye ugcina ixesha.
Iinkcukacha
50 Preps
Amacandelo ekhithi
| Isithinteli ST1 |
| Isithinteli ST2 |
| Umgca Acrylamide |
| Isithinteli PW |
| Buffer WB |
| Isithinteli EB |
| I-Foregene Protease |
| Ikholamu yeDNA-Kuphela |
| Imiyalelo |
Iimpawu&izinto eziluncedo
-Akukho ungcoliseko lwe-RNase: IKholamu ye-DNA-Kuphela enikezelwe yikiti yenza kube lula ukususa i-RNA kwi-DNA ye-genomic ngaphandle kokongeza i-RNase ngexesha lovavanyo, ukuthintela ilabhorethri ukuba ingangcoliswa yi-RNase yangaphandle.
-Isantya esikhawulezayo: I-Foregene Protease inomsebenzi ophezulu kuneeproteases ezifanayo, ukugaya iisampuli ngokukhawuleza;umsebenzi olula.
-Elula: I-centrifugation yenziwa kwiqondo lokushisa, kwaye akukho mfuneko ye-4 ° C ye-centrifugation ephantsi okanye i-ethanol precipitation ye-DNA.
-Ukhuseleko: Akukho kukhutshwa kwe-organic reagent efunekayo.
-Umgangatho ophezulu: I-DNA ye-genomic ekhutshweyo inamaqhekeza amakhulu, akukho RNA, akukho RNase, kunye nomxholo we-ion ophantsi kakhulu, onokuhlangabezana neemfuno zeemvavanyo ezahlukeneyo.
-Inkqubo ye-Micro-elution: Inokunyusa i-concentration ye-DNA ye-genomic, ekulungele ukufunyanwa kwe-downstream okanye uvavanyo.
Isicelo seKit
Ifanelekile ukuhlanjululwa kwe-DNA ye-genomic kwiisampuli ezilandelayo: i-buccal swabs, iKhadi le-FTA (amabala egazi).
Ugcino kunye nobomi beShelf
-Le khithi ingagcinwa kwiinyanga ezili-12 phantsi kweemeko ezomileyo kwindawo yokushisa (15-25 ° C);ukuba ifuna ukugcinwa ixesha elide, inokugcinwa kwi-2-8 ° C.
Qaphela: Ukuba igcinwe kwiqondo lobushushu eliphantsi, isisombululo sithande ukuba nemvula.Ngaphambi kokusetyenziswa, qiniseka ukuba ubeke isisombululo kwikhithi kwindawo yokushisa kwegumbi ixesha elithile.Ukuba kuyimfuneko, yitshise kwindawo yokuhlambela amanzi angama-37 ° C imizuzu eyi-10 ukutshabalalisa i-precipitate, kwaye udibanise ngaphambi kokusetyenziswa.
-Isisombululo seForegene Protease sinefomula ekhethekileyo, esebenzayo xa igcinwe kwindawo yokushisa ixesha elide (iinyanga ezi-3);umsebenzi wayo kunye nokuzinza kuya kuba ngcono xa igcinwe kwi-4 ° C, ngoko kuyacetyiswa ukuba igcinwe kwi-4 ° C, khumbula ukuba ungayigcini -20 ° C.
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.
Isincomo: Ixesha lokucubungula isampuli ye-Foregene Protease inokwandiswa ngokufanelekileyo okanye i-supernatant ingathathwa emva kwe-centrifugation kwi-12,000 rpm (~ 13,400 × g) kwi-5 min.
2. Ukusetyenziswa ngokugqithiseleyo kweesampuli zezicubu okanye izicubu ezinkulu.
Isincomo: Kungcono ukuba ungadluli kwi-1 Buccal 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 imiyalelo yesampulu ye-buccal swab kwisikhokelo sokusebenza, kwaye usule amaxesha amaninzi kangangoko ukuze iiseli ezaneleyo zincanyathiselwe 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 imiqathango yogcino lweForegene Protease okanye endaweni yayo ufake iForegene Protease entsha yokusabela kwi-enzymatic.
4. Ukugcinwa okungafanelekanga kwekiti okanye ixesha lokugcina ixesha elide kakhulu, okubangelwa ukungaphumeleli kwamanye amacandelo kwikiti.
Ingcebiso: Thenga ikiti entsha ye-Buccal swab DNA 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.
Isincomo: Yongeza i-65 °C yehla e-eluent eshushu ngaphambili ukuya embindini we-membrane ye-silicone kwaye ushiye kwindawo yobushushu bemizuzu emi-5 ukwandisa ukusebenza kakuhle kwe-elution.
I-DNA ye-genomic ephantsi yesivuno eyedwa
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 imiqathango yogcino lweForegene Protease okanye endaweni yayo ufake iForegene Protease entsha yokusabela kwi-enzymatic.
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 kwi-genomic DNA elution njengoko kufunwa kwimiyalelo, ubuncinane kungabikho ngaphantsi kwe-15 μl.
Ukucoceka okuphantsi kwe-DNA ye-genomic yodwa
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 acid isolation kits can remove RNA ngaphandle kongezelelo olongezelelweyo lweRNase, ngoko ke buccal Swab/FTA Card DNA Isolation Kit akufuneki ukuba yongeze iRNase;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.
