■ Kuyera kwambiri: Khungu la silika limachotsa zodetsa zambiri.
Kugwiritsa ntchito kwambiri: Oyenera magazi anticoagulant (EDTA, heparin etc.), chovala chofufumitsa komanso magazi kuundana.
■ Kugwira ntchito mwachangu: Genomic DNA itha kupezeka mkati mwa ola limodzi.
Type: Spin column based
Zitsanzo ndi zoyambira: 100-1000 μl magazi athunthu ochokera kwa mammalian, 5-20 μl magazi athunthu kuchokera ku mbalame kapena amphibian, 200-1000 μl magazi
Cholowera: Genomic DNA.
Zokolola: 3-40 μg
Nthawi yogwiritsira ntchito: ~ 1 ora
Kugwiritsa ntchito kumunsi: Kuletsa kwa michere ya enzyme, kusanthula kwa PCR, kumwera kwa blot, chip hybridization, ntchito yomanga laibulale ya NGS, ndi zina zambiri.
1. Kuti muchotse zotsalira za RNA, yankho la RNase A (100 mg / ml) liyenera kukhala lokonzekera.
2. Chogulitsachi chidalembetsedwa m'gulu la 1 mu m'galasireagent yodziwika ndi Beijing FDA. Chonde nditumizireni ku TIANGEN ngati mukufuna kudziwa zambiri.
Zogulitsa zonse zimatha kusinthidwa kukhala ODM / OEM. Zambiri,chonde dinani Makonda Service (ODM / OEM)
Genomic DNA idachotsedwa m'magazi pogwiritsa ntchito TIANamp Blood DNA Kit. M: Chizindikiro cha TIANGEN D15000. Zitsanzo: 200 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. Povala chovala chamtundu wa buffy, 2 μl of 100 μl eluates adakwezedwa; kwa enawo, 4 μl of 100 μl eluates adakwezedwa pamsewu. Zotsatira zoyesa: TIANamp Blood DNA Kit itha kugwiritsidwa ntchito kwambiri mu kuyeretsa kwapamwamba kwambiri kwa DNA kuchokera ku maanticoagulants aanthu okhala ndi mankhwala osiyanasiyana komanso magazi amitundu yosiyanasiyana. |
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Kuyerekeza kwa genomic DNA yochokera ku 200 μl anticoagulation ya anthu ndi EDTA pogwiritsa ntchito TIANamp Blood DNA Kit ndi zinthu zofananira kuchokera kwa omwe amapereka. M: Chizindikiro cha TIANGEN D15000. 4 μl ya 100 μl eluates adakwezedwa pamsewu. Zotsatira zoyesera: Yerekezerani ndi ogulitsa ena, TIANamp Blood DNA Kit itha kupeza zokolola zambiri zamagazi gDNA. |
A-1 Kuchuluka kwa maselo kapena kachilombo koyambitsa nyereredwe -Pinditsani 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. Ethanol imatha kuchotsedwa poika pakati gawo la spin kwa 3-5 min, kenako ndikuyika firiji 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 chopera kwambiri nayitrogeni wamadzi, kupezanso chinyezi, kapena kuchuluka kwachitsanzo.
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 ..