■ Zosintha zoyambira poyambira: 2-400 μg DNA yoyera kwambiri imatha kutengedwa kuchokera ku 0.1-20 ml ya mitundu ingapo yamagazi.
■ Otetezeka komanso odalirika: Palibe kuipitsidwa kwa zinthu zosungunulira zinthu monga phenol / chloroform.
Yoyenera kumagwiritsa ntchito pafupipafupi, monga choletsa chimbudzi, PCR, kumanga laibulale, Southern blot (chip hybridization of high-throughput sequence), ndi zina zambiri.
Zogulitsa zonse zimatha kusinthidwa kukhala ODM / OEM. Zambiri,chonde dinani Makonda Service (ODM / OEM)
Kuyeretsa kwa genomic DNA kuchokera ku 300 μl, 5 ml ndi 10 ml magazi a anticoagulation magazi ndi EDTA pogwiritsa ntchito RelaxGene Blood DNA System. 3 μl ya 200 μl, 500 μl ndi 1 ml eluates motsatizana adayikidwa pamseu uliwonse. M: Chizindikiro cha TIANGEN D15000. Zotsatira zoyesa: 0.1-20 ml yamagazi imatha kutulutsidwa ndi RelaxGene Blood DNA System yokhala ndi mzere wabwino. |
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Kuyerekeza kwa genomic DNA yotengedwa kuchokera ku 300 μl anticoagulation ya anthu ndi EDTA pogwiritsa ntchito RelaxGene Blood DNA System ndi zinthu zofananira kuchokera kwa omwe amapereka. M: Chizindikiro cha TIANGEN D15000. 3 μl ya 200 μl eluates adakwezedwa pamsewu. Zotsatira zoyesa: Yerekezerani ndi ogulitsa ena, RelaxGene Blood DNA System imatha kupeza zokolola zambiri, kuyera komanso kuphatikiza kwa magazi gDNA. |
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Genomic DNA idachotsedwa m'magazi pogwiritsa ntchito RelaxGene Blood DNA System. Zitsanzo: 300 μl magazi anticoagulation magazi ndi EDTA, EDTA / NaF, heparin ndi sodium citrate; buffy kuchokera 2 ml magazi; 10 μl magazi a nkhuku ndi magazi a 200 μl magazi. 3 μl ya 200 μl eluates adakwezedwa pamsewu. M: Chizindikiro cha TIANGEN D15000. Zotsatira zoyesera: RelaxGene Blood DNA System itha kugwiritsidwa ntchito kwambiri mu kuyeretsa kwapamwamba kwambiri kwa maantibayotiki opangidwa ndimankhwala osiyanasiyana komanso magazi amitundu yosiyanasiyana. |
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Zina Zotengera |
A-1 Kuchuluka kwa maselo kapena kachilombo koyambitsa nyereredwe yoyamba - Limbikitsani kuchuluka kwa ma cell kapena ma virus.
A-2 Kuperewera kwa sampuli -Zitsanzozo sizinasakanikirane bwino ndi chosungira cha lysis. Amalangizidwa kuti azisakanikirana bwino ndi kutentha kwa thupi maulendo 1-2. —Maselo osakwanira omwe amabwera chifukwa cha kuchepa kwa proteinase K. —Wosakwanira cell lysis kapena kuwonongeka kwa mapuloteni chifukwa chosakwanira nthawi yosamba. Amalangizidwa kuti azidula zidutswazo tizidutswa tating'ono ndikuwonjezera nthawi yosamba kuti achotse zotsala zonse mu lysate.
A-3 Kusakwanira kwa DNA kutsatsa. - Palibe ethanol kapena peresenti yotsika m'malo mwa 100% ethanol yomwe idawonjezeredwa lysate isanatumizidwe ku gawo loyambira.
A-4 Mtengo wa pH wa elution buffer ndiwotsika kwambiri. -Sinthani pH kuti ikhale pakati pa 8.0-8.3.
Mowa wotsalira mumtambo.
—Pali zotsalira zotsukira PW mumlengalenga. Mowa ungachotsedwe poyika gawo loyenda la 3-5 min, kenako ndikuyika kutentha kapena 50 ℃ incubator ya 1-2 min.
A-1 Chitsanzocho sichatsopano. —Tengani chitsanzo chabwino cha DNA kuti muwone ngati DNA yomwe ili mchitsanzocho yawonongeka.
A-2 Chithandizo chisanachitike. —Kumachitika chifukwa cha utsi wambiri wa nayitrogeni, kupezanso chinyezi, kapena kuchuluka kwa chitsanzocho.
Zoyesererazi ziyenera kusiyanasiyana pamitundu yosiyanasiyana. Pazitsamba, onetsetsani kuti mukugaya madzi asafe. Zitsanzo za nyama, homogenate kapena sungani bwino madzi asafe. Pazitsanzo zokhala ndi makoma osavuta kuswa, monga mabakiteriya a G + ndi yisiti, amalimbikitsidwa kugwiritsa ntchito lysozyme, lyticase kapena njira zamakina kuti athyole makoma am'maselo.
4992201/4992202 Plant Genomic DNA Kit imagwiritsa ntchito njira yozikika yomwe imafunikira chloroform kuti izichotse. Makamaka ndi oyenera mitundu yazomera zosiyanasiyana, komanso ufa wouma wouma. Hi-DNAsecure Plant Kit imapangidwanso m'mbali, koma osafunikira kuchotsedwa kwa phenol / chloroform, kuti ikhale yotetezeka komanso yopanda poizoni. Ndioyenera kumera ndi polysaccharides komanso polyphenol. 4992709/4992710 DNAquick Plant System imagwiritsa ntchito njira yopangira madzi. Kuchotsa kwa phenol / chloroform sikufunikanso. Njira yodziyeretsera ndiyosavuta komanso yachangu yopanda malire pazitsanzo zoyambira, kotero ogwiritsa ntchito amatha kusintha ndalamazo mosinthasintha malinga ndi zoyeserera. Kukula kwakukulu kwa zidutswa za gDNA kumatha kupezeka ndi zokolola zambiri.
Kutulutsa magazi kwamagazi a DNA kumatha kuchitidwa pogwiritsa ntchito ma reagents omwe amapezeka mu zida ziwirizi posintha ndondomekoyi pamalangizo apadera otulutsa magazi a DNA. Kope lofewa la pulogalamu yamagazi yamagazi ya DNA limatha kuperekedwa mukapempha.
Yimitsani zitsanzo zatsopano ndi 1 ml PBS, saline wamba kapena buff buff. Kwathunthu homogenize nyemba ndi homogenizer ndi kusonkhanitsa precipitate pansi pa chubu ndi centrifuging. Tayani supernatant, ndikuyambiranso kuyambiranso ndi 200 μl buffer GA. Kuyeretsa kwa DNA kungachitike potsatira malangizo.
Poyeretsa gDNA mu plasma, seramu ndi madzi madzimadzi, TIANamp Micro DNA Kit ikulimbikitsidwa. Poyeretsa kachilombo ka HIV kuchokera ku zitsanzo za seramu / plasma, TIANamp Virus DNA / RNA Kit ikulimbikitsidwa. Poyeretsa bakiteriya gDNA kuchokera ku ma seramu ndi zitsanzo za plasma, TIANamp Bacteria DNA Kit ikulimbikitsidwa (lysozyme iyenera kuphatikizidwa ndi bakiteriya wabwino). Mwa zitsanzo za malovu, Hi-Swab DNA Kit ndi TIANamp Bacteria DNA Kit amalimbikitsidwa.
Chida chodzikongoletsera cha DNAsec kapena DNAquick Plant System ndikulimbikitsidwa pakuchotsa mafangasi a genome. Pochotsa yisiti, TIANamp Yisiti ya Kit ikulimbikitsidwa (lyticase iyenera kudzikonzekeretsa).
Chiyambireni kukhazikitsidwa, fakitole yathu yakhala ikupanga zinthu zoyambirira motsatira mfundoyo
yaubwino woyamba. Zogulitsa zathu zapeza mbiri yabwino pamsika ndikudzidalira pakati pa makasitomala akale ndi akale ..