Hoʻomaka ka qPCR Premix me-UNG

Helu popoki: HCB5071E

ʻO Superstart qPCR Premix plus-UNG kahi reagent kūikawā i hoʻolālā ʻia no ka Real Time PCR qualitative a me quantitative reactions me ka hoʻohana ʻana i ka ʻimi noiʻi, hoʻomohala kūikawā no nā kaʻina lyophilization.Loaʻa iā ia kahi enzyme hoʻomaka wela Hotstart Taq plus (DG), nona kāna hana enzyme Taq i hoʻopaʻa ʻia i ka lumi wela, me ka pale ʻana i ka hoʻonui ʻole kikoʻī i hoʻokumu ʻia e ka annealing non-specific a i ʻole ka hoʻokumu ʻana o ka dimer primer ma lalo o nā kūlana haʻahaʻa haʻahaʻa, pēlā e hoʻomaikaʻi ai. ke kikoo o ka hoonui ana.Ke hoʻohana nei kēia reagent i kahi pahu kūʻokoʻa qPCR i hoʻopaʻa ʻia a me ka ʻōnaehana anti-contamination UNG/dUTP e hoʻokō i ka hoʻomaka wela wikiwiki, hoʻomaikaʻi nui i ka pono a me ka naʻau o nā pane qPCR.Hiki iā ia ke loaʻa i nā pihi maʻamau maikaʻi i nā wahi ākea ākea a hoʻokō pololei i ka quantitation, me ka pale ʻana i ka hoʻonui maikaʻi hewa ʻole i hana ʻia e nā huahana PCR koena a i ʻole aerosol contamination.Ua kūpono kēia reagent me nā mea hana PCR nui fluorescent mai nā mea hana e like me Applied Biosystems, Eppendorf, Bio-Rad a me Roche etc., a hōʻike i ka paʻa maikaʻi ma ke ʻano lyophilized.

2. He ʻokoʻa ka helu kope o nā genes i loaʻa i loko o nā ʻano mamana like ʻole;inā pono, hiki ke hana ʻia ka dilution gradient e hoʻoholo ai i ka nui o ka hoʻohui ʻana i ka template.

3. Hiki ke lyophilized kēia pūnaewele;i ka wā e hoʻohana ai nā mea kūʻai aku i kēia ʻōnaehana me ka ʻole o nā koi maloʻo maloʻo, hiki ke hoʻohui ʻia ka 4 × lyoprotectant; inā loaʻa nā huahana maloʻo maloʻo e koi ʻia, i ka wā o ka wai reagents e hōʻoia i ka hana huahana, pono ia e hoʻohui i ka 4 × lyoprotectant e hōʻoia i ka kūlike me nā ʻōnaehana lyophilized. a me nā hopena.

2. E hoʻohui i ka mamana nucleic acid i ka pauka lyophilized a hoʻohui i ka wai a hiki i 25µL;

3. E hui maikaʻi me ka centrifugation a holo ma ka mīkini.

 Mana maikaʻi:

1. Ka ho'āʻo hana: sensitivity, specificity, reproducibility o qPCR.

2. ʻAʻohe hana nuclease exogenous, ʻaʻohe exogenous endo/exonuclease contamination.

ʻIkepili ʻenehana:

1. Hoʻohana ʻo Superstart qPCR Premix plus-UNG i kahi enzyme hoʻomaka wela hou e hiki ai i ka hoʻomaka wela wikiwiki i loko o 1 ~ 5 mau minuke;ma o ka hoʻokumu ʻana i ka pale kūikawā kūpono ia no nā hopena PCR nui fluorescent quantitative.

2. He kiʻekiʻe kikoʻī e hoʻomaikaʻi nui i ka naʻau o ka fluorescence quantitative PCR palena ʻike, hana amplification curves normalization, fluorescence waiwai loaʻa maopopo ka hoʻomaikaʻi ʻana i nā haʻahaʻa haʻahaʻa haʻahaʻa, kūpono e like me ke kiʻekiʻe sensitivity fluorescence quantitative PCR detection reagents.

3. No nā mea mua me ka mahana annealing haʻahaʻa a ʻoi aku paha ma mua o nā ʻāpana 200bp, ʻōlelo ʻia ke ʻano 3-step.

4. ʻOkoʻa ka maikaʻi o ka hoʻohana ʻana o ka dUTP a me ka naʻau i ka enzyme UNG no nā ʻano genes ʻokoʻa, no laila, inā hoʻohana ʻia ka ʻōnaehana UNG i ka emi ʻana o ka ʻike ʻike, pono e hoʻoponopono ʻia ka ʻōnaehana hopena.Inā makemake ʻia ke kākoʻo ʻenehana e ʻoluʻolu e kelepona i kā mākou hui.

5. E hoʻohana i nā wahi i hoʻolaʻa ʻia a me nā pipette ma mua a ma hope o ka hoʻonui ʻana, e hoʻokomo i nā mīkina lima i ka wā e hana ai a hoʻololi pinepine iā lākou;mai wehe i ka paipu hopena ma hope o ka pau ʻana o ka PCR e hōʻemi i ka haumia o nā laʻana e nā huahana PCR.