Zebra Fish Direct PCR Kit Direct PCR Lysis Reagent(zebrafish)
ʻO ka wehewehe ʻana o ka pahu:
E hoʻohana pololei i ka zebrafish lysate ma ke ʻano he laʻana no ka hoʻonui ʻana i ka PCR, me ka ʻole o ka unuhi ʻokoʻa o ka DNA, wikiwiki a kiʻekiʻe.
◮ʻAʻole pono ka hoʻomaʻemaʻe DNA hoʻopau manawa a me ka pipiʻi.
◮He liʻiliʻi ke koi hāpana, lawe wale i hoʻokahi hua.
◮ʻAʻole koi ʻia nā hana kūikawā e like me ka wili a me ka ʻoki ʻana, a maʻalahi ka hana.
◮ʻO ka ʻōnaehana PCR i hoʻopaʻa ʻia e hiki ai i ka PCR ke loaʻa ka kikoʻī kiʻekiʻe aʻe a ʻoi aku ka ikaika o ka hoʻomanawanui i ka PCR reaction inhibitors.
Nā wehewehe
Ke hoʻohana nei kēia huahana 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 ka PCR reactions, no laila ua kūpono loa ia no ka hoʻāʻo ʻana o 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.
2×PCR maʻalahiTMHiki i ka Mix ke hoʻomanawanui ikaika i ka PCR reaction inhibitors, a hiki ke hoʻohana i ka lysate o ka hāpana e hoʻāʻo ʻia ma ke ʻano he laʻana no ka hoʻonui pono a kikoʻī.Aia kēia reagent i ka ForegeneD-Taq DNAPolymerase, dNTPs, MgCl2, pale pale, PCR optimizer a stabilizer.Hoʻohana pū ʻia me ka lysis buffer hiki ke ʻike maʻalahi i nā laʻana, a loaʻa iā ia nā hiʻohiʻona o ka naʻau kiʻekiʻe, ka kikoʻī ikaika, a me ke kūpaʻa maikaʻi.
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 maʻalahiTMHuikau |
| 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.
■ 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.
Ka holo 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, inā hoʻohana pinepine ʻia, hiki ke mālama ʻia ma 4 ℃ no ka mālama pōkole (hoʻohana i loko o 10 mau lā).
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