■ Njira yonseyi imatha kumaliza ola limodzi.
■ Sikofunika xylene kapena zinthu zina zosungunulira zoopsa.
■ DNA yoyera kwambiri itha kupangika popanda chimbudzi cha proteinase K usiku umodzi.
■ PCR ndi RT-qPCR
■ SNP ndi STR kusanthula majini
■ Kafukufuku wa Pharmacogenomics
Zogulitsa zonse zimatha kusinthidwa kukhala ODM / OEM. Zambiri,chonde dinani Makonda Service (ODM / OEM)
DNA ya genomic yotulutsidwa ndi TIANquick FFPE DNA Kit ndi zinthu zofunikira kuchokera ku Supplier A zidakulitsidwa kukhala 190 bp, 300 bp, 750 bp zidutswa. Zotsatira zake zikuwonetsa kuti kukweza kwamphamvu kwa PCR kwa gDNA yotulutsidwa ndi TIANquick FFPE DNA Kit ndi Supplier A zinali zofanana.
Kuyamba zakuthupi: Mbewa ya chiwindi FFPE zitsanzo (zidutswa 5, 10 μm / chidutswa)
Ndondomeko: Tsatirani malangizo a chinthu chilichonse.
Kutsegula gel:
gDNA idasangalatsidwa ndi 50 μl elution buffer ndipo idakwezedwa kukhala 190 bp, 300 bp ndi 750 bp. 5μl ya 20 μl zopangidwa ndi PCR zidakwezedwa pamsewu umodzi pa 1% agarose gel. Electrophoresis inkachitika pa 6 V / cm kwa mphindi 20.
M: Chizindikiro cha TIANGEN D2000;
T: TIANquick FFPE DNA zida;
A: Zofanana ndi izi kuchokera kwa Wogulitsa A.
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 ..