ʻO Zebra Fish Direct PCR Kit-UNG Direct PCR Lysis Reagent(zebrafish)
Nā wehewehe
Ke hoʻohana nei kēia pahu i kahi ʻōnaehana lysis buffer kūʻokoʻa e hoʻokuʻu koke i ka DNA genomic mai ka iʻa zebrafish a me nā ʻiʻo iʻa wai hou, nā huelo a i ʻole nā hua hua iʻa no nā hopena PCR, no laila kūpono loa ia no ka hoʻāʻo ʻana i ka genetic nui.Hoʻopau ʻia ke kaʻina hana o ka hoʻokuʻu ʻana i ka DNA genomic mai ka lysis buffer i loko o 10-30 mau minuke ma 65 ° C.ʻAʻole koi ʻia nā kaʻina hana ʻē aʻe e like me ka protein a me ka wehe ʻana i ka RNA, a hiki ke hoʻohana ʻia ka trace DNA i hoʻokuʻu ʻia ma ke ʻano he kumu hoʻohālike no ka hopena PCR.
Hoʻohana ʻo 2× PCR EasyTM Mix (UNG) i ka dUTP ma kahi o ka dTTP ma ke kumu o 2× PCR EasyTM Mix, a hoʻohui i ka enzyme UNG (Uracil-N-glycosylase) hiki ke hoʻohaʻahaʻa i ka template i loaʻa ka dUTP i ka manawa like.Ma mua o ka hopena PCR, hoʻohana ʻia ka enzyme UNG e hoʻohaʻahaʻa i ka huahana PCR i loaʻa ka uracil.ʻAʻole loaʻa ka hopena o ka enzyme UNG i ka template i loaʻa ʻole ka uracil, no laila e hōʻoiaʻiʻo ai i ka kikoʻī a me ka pololei o ka amplification a me ka pale ʻana i ka hiki ke hoʻāʻo genetic nui.Ua hoʻopōʻino ʻia nā huahana PCR.
ʻO D-Taq DNA polymerase kahi DNA polymerase i hoʻomohala kūikawā ʻia e Foregene no nā hopena PCR pololei.Hiki i ka D-Taq DNA polymerase ke hoʻomanawanui i ka PCR reaction inhibitors, a hiki ke hoʻonui i ka nui o ka DNA i loko o nā ʻōnaehana hoʻololi paʻakikī, a hiki i ka wikiwiki hoʻonui ke hiki i 2Kb/min, kūpono loa ia no ka pane pololei PCR.
Nā kikoʻī
50×20µl rxns, 200×20µl rxns, 500×20µl rxns, 2000×20µl rxns
Nā ʻāpana pahu
Mahele I | Hoʻopaʻa FP |
Foregene Protease | |
6× DNA Hoʻouka Pahu | |
Mahele II | 2×PCR maʻalahiTMHui (UNG) |
Nā kuhikuhi |
Nā hiʻohiʻona & pono
■ ʻAʻole pono ka hoʻomaʻemaʻe DNA hoʻopau manawa a me ka pipiʻi.
■ He liʻiliʻi ke koi hoʻohālike, hiki ke hoʻāʻo ʻia e like me 1mg iʻa caudal fin a i ʻole 10 hua iʻa.
■ ʻAʻole koi ʻia nā hana kūikawā e like me ka wili a me ka ʻoki ʻana, a maʻalahi ka hana.
■ Pūnaehana PCR Anti-pollution 2× PCR EasyTM Mix (UNG), hiki ke hoʻopau pono i ka haumia i hana ʻia e nā huahana PCR a hōʻoia i ka kikoʻī a me ka pololei o ka amplification.
■ ʻOi aku ka maikaʻi o ka PCR ʻoi aku ka maikaʻi o ka hoʻomanawanui ʻana i ka PCR reaction inhibitors.
Nā palena kime
Hoʻohana: ka iʻa Zebra a me nā iʻa wai hou ʻē aʻe.
Hoʻokuʻu ʻia ka DNA mai ka laʻana lysis: hoʻohana wale ʻia ma ke ʻano he PCR template.
Hiki ke hoʻohana ʻia ke kit no nā kumu penei: ʻike i ka transgenosis, genotyping holoholona, etc.
Kaʻina hana
Ka mālama ʻana a me ka mālama ʻana
Pono e mālama ʻia ka ʻāpana I o kēia pahu ma 2-8 ℃.
Hiki ke mālama ʻia ka reagent Buffer FP no 12 mahina ma lalo o nā kūlana maloʻo;hiki ke mālama ʻia ma 2-8 ℃ no ka mālama lōʻihi.
v Loaʻa i ka Reagent Foregene Protease kahi kumu kūʻokoʻa.No ka hōʻoia ʻana i kāna hana a paʻa, e ʻoluʻolu e mālama iā ia ma 4 ℃ no 12 mahina.
vReagent 6×DNA Loading Buffer hiki ke mālama ʻia ma 4 ℃ a i ʻole -20 ℃ no ka manawa lōʻihi.
Pono e mālama ʻia ka ʻāpana II o kēia pahu ma -20 ℃.
vReagent 2×PCR EasyTM Mix(UNG), inā hoʻohana pinepine ʻia, hiki ke mālama ʻia ma 4 ℃ no ka mālama manawa pōkole (hoʻohana i loko o 10 mau lā).