I-Bacterial DNA Isolation Kit I-Bacterial Genomic DNA Extraction Iikhithi zokuCoca
Inkcazelo yeKit:
Ngokukhawuleza uhlambulule i-DNA ye-genomic ephezulu evela kwiibhaktheriya kwisigaba sokukhula kwe-logarithmic.
◮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 inika indlela ekhawulezayo nelula yokukhupha i-DNA ye-genomic kwiinkcubeko zebhaktheriya (i-Gram-negative kunye ne-Gram-positive) kwimithombo eyahlukeneyo;inokuqhuba ngaphantsi kwe-3ml yeenkcubeko zebhaktheriya kwisigaba sokukhula kwe-logarithmic (i-1 × 109 ibhaktheriya).Ikiti ihambelana nokusebenza okuphezulu kwe-Foregene Protease, evumela i-15-50μg ye-DNA ye-genomic ephezulu ukuba ikhutshwe kwiyure ye-1.Ukongeza, ikiti inokukhupha izinto zofuzo ngaphandle kwe-genome, njenge-plasmids, iCosmid, i-BAC, njl.
I-DNA-kuphela kwe-silica gel matrix impahla esetyenziswe kwikholamu ye-spin yinto entsha ekhethekileyo yenkampani, enokuthi ibhengeze ngokufanelekileyo kwaye ibhengeze i-DNA.Umthamo omkhulu we-adsorption we-DNA yi-80μg.Inkqubo ekhethekileyo ye-buffer kunye ne-elution inokwandisa ukususwa kwe-RNA, iiprotheni ezingcolileyo, ii-ion kunye nezinye izinto eziphilayo kwiiseli.Iqhekeza le-DNA ye-genomic likhulu, liphezulu, lizinzile kwaye linomgangatho othembekileyo, kwaye ubukhulu beqhekeza le-DNA buzinzile malunga ne-23kb.
Iinkcukacha
50 Preps, 100 Preps, 250 Preps
Amacandelo ekhithi
| Isithinteli ML1 |
| Isithinteli ML2 |
| Isithinteli PW |
| Buffer WB |
| Isithinteli EB |
| I-Foregene Protease |
| I-Lysozyme |
| Isithinteli TE |
| Ikholamu yeDNA-Kuphela |
| Imiyalelo |
Iimpawu&izinto eziluncedo
-Akukho ungcoliseko lwe-RNase: Akukho mfuneko yokongeza i-RNase ukususa i-RNA kwi-DNA ye-genomic, ukuphepha i-laboratory ukuba ingangcoliswa yi-RNase yangaphandle.
-Isantya esikhawulezayo: I-Foregene Protease inomsebenzi ophezulu kuneeproteases ezifanayo kwaye inokwetyisa iisampuli zebhaktheriya ezaphukileyo ngexesha elifutshane.
-Uncedo: 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: Iziqwenga ze-DNA ze-genomic ezikhutshwe zikhulu, akukho RNA, akukho RNase, kunye nomxholo we-ion ophantsi kakhulu, onokuhlangabezana neemfuno zeemvavanyo ezahlukeneyo.
Isicelo seKit
Le khithi ilungele ukuhlanjululwa kwe-DNA ye-genomic kwezi sampuli zilandelayo: ibhaktheriya ye-Gram-negative kunye ne-Gram-positive bacteria kwisigaba sokukhula kwe-logarithmic.
Ukuhamba komsebenzi
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);xa igcinwe kwi-4 ° C, umsebenzi wayo kunye nokuzinza kuya kuba ngcono, ngoko kuyacetyiswa ukuba uyigcine kwi-4 ° C, khumbula ukuba ungayigcini -20 ° C.
-Lysozyme ngumgubo owomileyo, osebenzayo xa ugcinwe kwi-2-8 ° C ixesha elide (iinyanga ezi-3).Ukugcina ixesha elide, nceda uyigcine ku -20°C.
