■ Isamba esiguqukayo semali yokuqala: 2-400 μg ubumsulwa obuphezulu be-DNA bungakhishwa ku-0.1-20 ml wamasampuli egazi ahlukahlukene.
■ Kuphephile futhi kunokwethenjelwa: Akukho ukungcoliswa kwama-solvents angokwemvelo njenge-phenol / chloroform.
Ifanele izinhlelo ezejwayelekile ezahlukahlukene, njengokuvinjelwa kokugaywa, i-PCR, ukwakhiwa kwelabhulali, iSouthern blot (i-chip hybridization yokulandelana okuphezulu), njll.
Yonke imikhiqizo ingenziwa ngezifiso i-ODM / OEM. Ngemininingwane,sicela uchofoze i-Customized Service (ODM / OEM)
Ukuhlanzwa kwe-genomic DNA kusuka ku-300 μl, 5 ml no-10 ml we-anticoagulation yegazi nge-EDTA kusetshenziswa iRelaxGene Blood DNA System. I-3 μl ye-200 μl, i-500 μl ne-1 ml eluates ngokulandelana yalayishwa kulayini ngamunye. M: Umaka we-TIANGEN D15000. Imiphumela yokuhlola: u-0.1-20 ml wegazi angakhishwa yi-RelaxGene Blood DNA System enobubanzi obuhle bomugqa. |
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Ukuqhathanisa i-genomic DNA ekhishwe ku-300 μl i-anticoagulation yabantu ne-EDTA kusetshenziswa iRelaxGene Blood DNA System nemikhiqizo efanayo evela kwabanye abahlinzeki. M: Umaka we-TIANGEN D15000. I-3 μl yama-200 μl eluates alayishwe kulayini ngamunye. Imiphumela yokuhlola: Qhathanisa nabanye abahlinzeki, iRelaxGene Blood DNA System ingathola isivuno esiphezulu, ubumsulwa nokuhlanganiswa kwegazi gDNA. |
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IGenomic DNA ikhishwe kumasampula egazi kusetshenziswa iRelaxGene Blood DNA System. Amasampula: I-300 μl igazi le-anticoagulation yabantu nge-EDTA, EDTA / NaF, heparin ne-sodium citrate; ijazi le-buffy kusuka egazini le-2 ml; Igazi lenkukhu elingu-10 μl negazi legundane elingu-200 μl. I-3 μl yama-200 μl eluates alayishwe kulayini ngamunye. M: Umaka we-TIANGEN D15000. Imiphumela yokuhlola: I-RelaxGene Blood DNA System ingasetshenziswa kabanzi ekuhlanzweni kwekhwalithi ephezulu ye-DNA yama-anticoagulants womuntu ngemithi ehlukene kanye negazi elivela ezinhlotsheni ezahlukahlukene. |
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Eminye Imininingwane Yesethenjwa |
A-1 Ukuhlushwa okuphansi kwamaseli noma igciwane kusampula yokuqala -Nothisa ukuhlushwa kwamaseli noma amagciwane.
I-A-2 I-lysis enganele yamasampuli-Amasampula awakaxutshwa kahle ne-lysis buffer. Kuphakanyiswa ukuthi uhlanganise kahle nge-pulse-vortexing izikhathi eziyi-1-2. —I-lysis enganele yeseli ebangelwe ukwehla komsebenzi we-proteinase K. —Ukungalingani kwe-cell lysis noma ukwehla kwamaprotheni ngenxa yesikhathi esanele sokugeza. Kuphakanyiswa ukusika izicubu zibe yizicucu ezincane bese unweba isikhathi sokugeza ukuze ususe yonke insalela ku-lysate.
I-A-3 Ukukhangisa okwanele kwe-DNA. -Ayikho i-ethanol noma amaphesenti aphansi esikhundleni se-100% ethanol engezwe ngaphambi kokuthi i-lysate idluliselwe kukholamu ye-spin.
A-4 Inani le-pH le-elution buffer liphansi kakhulu. - Lungisa i-pH iye phakathi kuka-8.0-8.3.
I-ethanol eyinsalela ku-eluent.
—Kunensalela yokuwasha i-PW esikhaleni. I-ethanol ingasuswa ngokufaka phakathi ikholamu ye-spin ngemizuzu engu-3-5, bese ifakwa ekamelweni lokushisa noma ku-50 ℃ incubator ye-1-2 min.
A-1 Isampula akuyona entsha. —Khipha i-DNA eyisampula enhle njengokulawula ukunquma ukuthi ngabe i-DNA esampulweni yehle yini.
Ukwelashwa kwangaphambili okungalungile. —Kudalwa ukugaya okweqile kwe-nitrogen, uketshezi olubuyisa umswakama, noma inani elikhulu kakhulu lesampula.
Ama-pretreatment kufanele ahluke kumasampula ahlukile. Ngamasampula ezitshalo, qiniseka ukuthi ugaya kahle i-nitrogen engamanzi. Ngamasampuli ezilwane, homogenate noma gaya kahle i-nitrogen engamanzi. Kumasampuli anezindonga zamaseli okunzima ukugqashuka, njengama-bacterium e-G + nemvubelo, kuphakanyiswa ukusebenzisa i-lysozyme, i-lyticase noma izindlela zemishini yokwephula izindonga zeseli.
I-4992201/4992202 iPlant Genomic DNA Kit isebenzisa indlela esekwe kukholamu edinga i-chloroform yokukhipha. Ilungele ikakhulukazi amasampula ezitshalo ezahlukahlukene, kanye ne-powder eyomile yesitshalo. IHi-DNAsecure Plant Kit nayo isekelwe kwikholomu, kepha asikho isidingo sokukhishwa kwe-phenol / chloroform, okuyenza iphephe futhi ingabi nobuthi. Ifanele izitshalo ezinama-polysaccharides aphezulu nokuqukethwe kwe-polyphenol. I-4992709/4992710 DNAquick Plant System isebenzisa indlela esekelwa uketshezi. Ukukhishwa kwePhenol / chloroform akudingeki futhi. Inqubo yokuhlanzwa ilula futhi iyashesha ngaphandle komkhawulo wamanani okuqala esampula, ngakho-ke abasebenzisi bangalungisa inani ngokuvumelana nezidingo zokuhlola. Usayizi omkhulu wezingcezu ze-gDNA ungatholakala ngesivuno esikhulu.
Ukukhishwa kwegazi kwe-DNA kwegazi kungenziwa kusetshenziswa ama-reagents ahlinzekwe kulawa makhithi amabili ngokumane uguqule umthetho olandelwayo uye emyalweni othize wokukhishwa kwegazi kwe-DNA. Ikhophi ethambile ye-clot yegazi ye-DNA protocol ingakhishwa uma ucela.
Misa okwesikhashana isampula elisha nge-1 ml PBS, usawoti ojwayelekile noma i-TE buffer. Homogenize Ngokuphelele isampula nge-homogenizer bese uqoqa imvula phansi kwephubhu nge-centrifuging. Lahla amandla angaphezu kwawemvelo, uphinde uvuselele ukugqashuka nge-200 μl buffer GA. Ukuhlanzwa kwe-DNA okulandelayo kungenziwa ngokuya ngemiyalo.
Ukuhlanzwa kwe-gDNA kumasampula e-plasma, serum kanye ne-fluid fluid, Kunconywa i-TIANamp Micro DNA Kit. Ukuhlanzwa kwegciwane i-gDNA kusuka kumasampula we-serum / plasma, Kunconywa i-TIANamp Virus DNA / RNA Kit. Ukuhlanzwa kwe-gDNA yebhaktheriya kusuka kumasampula we-serum ne-plasma, i-TIANamp Bacteria DNA Kit inconywa (i-lysozyme kufanele ifakwe ibhaktheriya elihle). Kumasampula amathe, i-Hi-Swab DNA Kit ne-TIANamp Bacteria DNA Kit kuyanconywa.
I-DNAsecure Plant Kit noma i-DNAquick Plant System inconyelwa ukukhishwa kwefungal genome. Ngokukhishwa kwemvubelo ye-genome, Kunconywa i-TIANamp Yeast DNA Kit (i-lyticase kufanele izilungiselele).
Kusukela yasungulwa, imboni yethu ibilokhu ithuthukisa imikhiqizo esezingeni lomhlaba yokuqala ngokunamathela esimisweni
wekhwalithi kuqala. Imikhiqizo yethu izuze idumela elihle kakhulu embonini futhi ithembekile phakathi kwamakhasimende amasha namadala ..