I-Plat DNA Isolation Kit Genomic Plant DNA yokuCoca iiKits Protocol
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Le khithi isebenzisa ikholamu ye-DNA kuphela enokuthi ibophe ngokukodwa i-DNA, i-Foregene protease kunye nenkqubo ye-buffer ekhethekileyo, eyenza lula kakhulu ukucocwa kwe-DNA ye-genomic yezityalo.I-DNA ye-genomic ephezulu inokufumaneka ngaphakathi kwemizuzu engama-30, ekhusela ukuthotywa kwe-DNA ye-genomic.
I-DNA-kuphela kwe-silica gel membrane esetyenziswe kwikholamu ye-spin yinto entsha ye-Foregene yezinto ezintsha, ezinokuthi zibophe ngokufanelekileyo kunye ngokuthe ngqo kwi-DNA, kwaye zandise ukususwa kwe-RNA, iiprotheni ezingcolileyo, i-ion, i-polysaccharides, i-polyphenols kunye nezinye izinto eziphilayo.
Amacandelo emveliso
| Buffer PL1, Buffer PL2 |
| I-Buffer PW, i-Buffer WB, i-Buffer EB |
| I-Foregene Protease |
| Ikholamu yeDNA-Kuphela |
| Imiyalelo |
Iimpawu&izinto eziluncedo
■ Akukho ngcoliseko lwe-RNase: IKholamu ye-DNA-Kuphela ebonelelwe yikiti yenza kube lula ukususa i-RNA kwi-DNA ye-genomic ngaphandle kwe-RNase eyongezelelweyo ngexesha lovavanyo, ukuthintela ilabhoratri ukuba ingangcoliswa yi-RNase yangaphandle.
■ Isantya esikhawulezayo: I-Foregene Protease inomsebenzi ophezulu kuneeprotease ezifanayo kwaye yetyise iisampuli zethishu ngokukhawuleza.
■ Lula: umsebenzi wokutsalwa kwe-genomic DNA unokugqitywa kwimizuzu engama-30.
■ Ukulungeleka: I-centrifugation yenziwa kwiqondo lobushushu begumbi, akukho 4℃ ubushushu obuphantsi becentrifugation okanye i-ethanol precipitation ye-DNA efunekayo.
■ Ukhuseleko: akukho sixhobo sokwenza izinto eziphilayo esifunekayo.
■ Umgangatho ophezulu: I-DNA ecociweyo ye-genomic inamaqhekeza amakhulu, ayikho i-RNA, ayikho i-RNase, kunye nomxholo we-ion ophantsi kakhulu, unokuhlangabezana neemfuno zemifuniselo eyahlukeneyo.
Isicelo seKit
Ilungele ukutsalwa kunye nokucocwa kwe-DNA ye-genomic ukusuka kwizicubu zezityalo ezitsha okanye ezikhenkcezisiweyo.
Ukuhamba komsebenzi
Umzobo
Ugcino kunye nobomi beShelf
Ikhithi ingagcinwa iinyanga ezili-12 kwiqondo lobushushu begumbi (15–25 ℃) kunye ne-2–8℃ ixesha elide.
Isisombululo se-Foregene Protease Plus sinefomula ekhethekileyo, esebenzayo xa igcinwe kwindawo yokushisa ixesha elide (iinyanga ezi-3);umsebenzi wayo kunye nokuzinza kuya kuba ngcono xa igcinwe4℃, ngoko kuyacetyiswa ukuba uyigcine ku-4℃, khumbula ukuba ungayigcini ku -20℃.
Isikhokelo sokuHlalutya ingxaki
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 esiphantsi okanye akukho DNA
Ngokuqhelekileyo kukho izinto ezininzi ezichaphazela isivuno se-DNA ye-genomic, kubandakanywa nomthombo wesampuli, iminyaka yesampuli, iimeko zokugcina isampuli, kunye nokusebenza.
I-Genomic DNA ayifumanekanga ngexesha lokukhutshwa
1. Iisampuli zezicubu azigcinwa ngokungafanelekanga okanye zigcinwe ixesha elide, okubangela ukuthotywa kwe-DNA ye-genomic.
Isincomo: Gcina iisampulu zethishu kwi-nitrogen engamanzi okanye -20°C;zama ukusebenzisa iisampulu ezisanda kuqokelelwa kwi-genomic DNA extraction.
2. Iisampulu encinci kakhulu inokubangela ukuba i-DNA ye-genomic ehambelanayo ingakhutshwa.
Ingcebiso: Kwiisampulu zethishu eziye zagcinwa ixesha elide okanye ezinokonakala okukhulu kwe-DNA ye-genomic, isixa seesampulu zezicubu zinokunyuswa ngokufanelekileyo ukuze kukhutshwe i-DNA ye-genomic.Isixa sesampuli sinokumiselwa ngokweemfuno ze-DNA, kodwa isampuli esitsha akufanele idlule i-100mg, kwaye isampuli eyomileyo akufanele idlule i-30mg.
3. Isampulu ayigutywa ngenitrogen engamanzi okanye ibekwe ixesha elide emva kwenitrogen engamanzi.
Isiphakamiso: Ngexesha lokukhutshwa kwe-DNA, isampuli kufuneka ihlatywe ngokupheleleyo kunye ne-nitrogen engamanzi ukuphula udonga lweseli;emva kokugaya, nceda udlulisele isampuli yempuphu kwi-PL1 efudumalayo kwi-65°C ngokukhawuleza (emva kokuba i-powder yomhlaba iyancibilika, i-DNA ye-genomic iya kuqala ukuthotywa ngokukhawuleza) .
4. Ukugcinwa okungafanelekanga kwe-Foregene Protease kubangela umsebenzi oncitshisiweyo okanye ongasebenziyo.
Ingcebiso: Qinisekisa imiqathango yokugcinwa kweForegene Protease okanye endaweni yayo ufake iForegene Protease entsha ye-enzymatic hydrolysis.
5. Ikhithi igcinwe ngokungafanelekanga okanye igcinwe ixesha elide kakhulu, ibangela ukuba amanye amacandelo ekhithi angaphumeleli.
Ingcebiso: Thenga isixhobo esitsha se-genomic DNA extraction kit ukwenzela imisebenzi enxulumeneyo.
6. Ukusetyenziswa ngokungafanelekanga kwekhithi.
Ingcebiso: Thenga i-Plant DNA Isolation Kit enikezelwe kwiisampulu zokukhutshwa kunye nokucocwa kwesityalo se-genomic DNA.
7. Isithinteli WB ngaphandle kokongeza aengenamanzi i-ethanol.
Ingcebiso: Qinisekisa ukuba wongeza umthamo ochanekileyo we-ethanol epheleleyo kwi-Buffer WB.
8. I-eluent ayizange ithontsizwe kwi-silica membrane ngokuchanekileyo.
Ingcebiso: Yongeza i-eluent eshushu ngaphambili kwi-65℃i-dropwise ukuya embindini we-silica gel inwebu, kwaye uyishiye kwiqondo lobushushu begumbi imizuzu emi-5 ukunyusa ukusebenza kakuhle kwe-elution.
Ukutsalwa ukufumana i-DNA ye-genomic ephantsi
1. Isampuli ayigcinwanga ngokungafanelekanga okanye igcinwe ixesha elide, okubangela ukuthotywa kwe-DNA ye-genomic.
Isincomo: Gcina iisampulu zethishu kwi -20℃;zama ukusebenzisa iisampulu zethishu ezisanda kuqokelelwa ukuze kutsalwe i-genomic DNA.
2. Ukuba inani leesampuli zezicubu lincinci kakhulu, i-DNA ye-genomic ekhutshweyo iya kuba ngaphantsi.
Ingcebiso: Ezinye iisampulu zezityalo zizityebi emanzini, njengezityalo zasemanzini ezifana ne-algae, njl., idosi inokongezwa ngokufanelekileyo okanye amanzi anokuphelelwa ngamanzi kancinci ngaphambi kotyando.
3. Iisampulu azizange zigatywe kakuhle ngenitrogen engamanzi okanye zishiywe kwindawo yobushushu begumbi ixesha elide emva kokusila.
Ingcebiso: Ulwelo lokusila initrogen kufuneka lwanele, kwaye isampulu yodonga lweseli kufuneka yaphulwe kangangoko;ngokukhawuleza emva kokusila, isampuli yomgubo kufuneka idluliselwe kwi-65℃ifudumeze iBuffer PL1 yenyathelo elilandelayo.
4. Ukungasebenzisi ikhithi echanekileyo.
Isincomo: Sebenzisa i-Plant DNA Isolation Kit ezinikeleyo ukuze ukhuphe kwaye uhlambulule i-genomic DNA yesityalo.
5. Ukugcinwa okungafanelekanga kwe-Foregene Protease kubangela umsebenzi oncitshisiweyo okanye ongasebenziyo.
Ingcebiso: Qinisekisa imiqathango yokugcinwa kweForegene Protease okanye endaweni yayo ufake iForegene Protease entsha ye-enzymatic hydrolysis.
6. Ingxaki ye-Eluent
Ingcebiso: Nceda usebenzise i-Buffer EB yolution;ukuba usebenzisa i-ddH2O okanye ezinye ii-eluents, qiniseka ukuba i-pH ye-eluent iphakathi kwe-7.0-8.5.
7. I-eluent ayivuthwanga ngokuchanekileyo
Ingcebiso: Nceda wongeze i-elution drop kumbindi we-silica inwebu kwaye uyishiye kwindawo yobushushu begumbi imizuzu emi-5 ukwandisa ukusebenza kakuhle kwe-lution.
8. Ivolumu ye-eluent incinci kakhulu
Ingcebiso: Nceda usebenzise i-eluent ye-genomic DNA elution ngokwemiyalelo, ubuncinane ingabi ngaphantsi kwe-100.μl.
I-DNA ye-genomic ekhutshwe ngokucoceka okuphantsi
Ukucoceka okuphantsi kwe-DNA ye-genomic kuya kubangela ukungaphumeleli okanye umphumo ombi weemvavanyo ezisezantsi, ezifana: i-enzyme ayinakunqunyulwa, kwaye i-fragment ye-gene ejoliswe kuyo ayinakufunyanwa yi-PCR.
1. Ukungcoliseka kweeprotheyini ezahlukeneyo, ukungcoliswa kwe-RNA.
Uhlalutyo: I-Buffer PW ayizange isetyenziswe ukuhlamba ikholamu;I-Buffer PW ayizange isetyenziswe ukuhlamba ikholamu kwisantya esichanekileyo se-centrifugation.
Isiphakamiso: zama ukuqinisekisa ukuba akukho mvula kwi-supernatant xa i-supernatant idlula kwikholamu;qiniseka ukuba uhlamba ikholamu yokucoca nge-Buffer PW ngokwemiyalelo, kwaye eli nyathelo alinako ukushiywa.
2. Ungcoliseko lwe-ion.
Uhlalutyo: Ikholamu yokuhlamba ye-Buffer WB ayifakwanga okanye ihlanjwe kube kanye kuphela, ibangele ungcoliseko lwe-ionic olushiyekileyo.
Ingcebiso: Qinisekisa ukuba uhlamba kabini nge-Buffer WB ngokwemiyalelo yokususa iiyoni ezishiyekileyo kangangoko kunokwenzeka.
3. Usulelo lweRNase.
Uhlalutyo: Exogenous RNase yongezwa kwi-Buffer;umsebenzi wokuhlamba ongalunganga kwi-Buffer PW uya kubangela intsalela ye-RNase kwaye ichaphazele imisebenzi yovavanyo ye-RNA esezantsi, efana ne-in vitro transcription.
Isiphakamiso: Iikiti ze-Foregene ze-nucleic acid extraction kits zinokususa i-RNA ngaphandle kwe-RNase eyongezelelweyo, kwaye zonke ii-reagents kwi-Plant DNA Isolation Kit ayifuni i-RNase;qiniseka ukuba uhlamba ikholamu yokucoca nge-Buffer PW ngokwemiyalelo, kwaye eli nyathelo alinako ukushiywa.
4. Iintsalela ze-Ethanol.
Uhlalutyo: Emva kokuhlamba ikholamu yokucoca nge-Buffer WB, akukho ityhubhu engenanto ye-centrifugation eyenziwa.
Isindululo: Landela imiyalelo yetyhubhu engenanto eyiyo ye-centrifugation.
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