2×Sensi Direct Premix-UNG (Probe qPCR)

Helu popoki: HCB5151A

Hoʻolālā ʻia ʻo SensiDirect Premix-UNG (Probe qPCR) e hana pololei i ka PCR mai nā laʻana me ka ʻole o ka unuhi ʻana o DNA a i ʻole ka hoʻomākaukau ʻana i ka laʻana.Aia kēia reagent me ka DNA polymerase wela, uracil DNA glycosylase (UNG), RNase Inhibitor, MgCl₂, dNTPs (me dUTP ma kahi o dTTP), a me nā stabilizers, no ka quantitative PCR (qPCR).Keia reagent preforms kiʻekiʻe inhibitor-hoʻomanawanui, a pela e hiki ke hoʻopili pololei i ka ʻike ʻana i nā laʻana e like me ka ʻāʻī ʻāʻī, kohu, anti-coagulated koko holoʻokoʻa, plasma, a me ka serum me ka ʻole o ka DNA extraction.Hoʻohana ka reagent i ka paʻa pono no ka qPCR me nā enzyme i hui pū ʻia o ka polymerase DNA anti-inhibitory a me ka enzyme UNG.No laila, hiki iā ia ke loaʻa ka hoʻonui maikaʻi ʻana o nā genes i hoʻopaʻa ʻia i nā laʻana i loaʻa nā inhibitors a keʻakeʻa i ka hoʻonui maikaʻi hewa ʻole i hana ʻia e ka PCR koena a me ka pollution aerosol.Ua kūpono kēia reagent me ka hapa nui o nā mea hana PCR fluorescent quantitative, e like me Applied Biosystems, Eppendorf, Bio-Rad, Roche a pēlā aku.

Nā ʻāpana

1. 50×SensiDirect Enzyme/UNG Mix

2. 2×SensiDirect Premix Buffer (dUTP)

Nā kūlana mālama

Pono e mālama ʻia nā ʻāpana āpau ma -20 ℃ no ka mālama lōʻihi a me 4 ℃ a hiki i 3 mahina.E ʻoluʻolu e hui pono ma hope o ka hoʻoheheʻe ʻana a me ka centrifuge ma mua o ka hoʻohana ʻana.Hōʻalo i ka hoʻoheheʻe pinepine ʻana.

Kūkākūkā Kaʻikala

Na kuhikuhi paipu

1. ʻO ka manaʻo hope loa o ka primer he 0.2μM.No nā hualoaʻa maikaʻi aʻe, hiki ke hoʻopaʻa ʻia ke kumu kumu i loko o ka laulā o 0.2-1μM.

2. ʻO ka maʻamau, hiki ke hoʻonui ʻia ka ʻimi noiʻi i loko o ka laulā o 0.1-0.3μM.ʻO ka manaʻo kūpono o ka probe e pili ana i ka mea hana PCR amplification manawa maoli, ke ʻano o ka probe, a me ke ʻano o ka mea hōʻailona fluorescent.E ʻoluʻolu e nānā i ka manual mea hana a i ʻole nā ​​koi kikoʻī o kēlā me kēia ʻimi fluorescent.

3. Aia nā ʻano laʻana like ʻole i nā ʻano like ʻole a me nā maʻiʻo o ka mea hoʻopaneʻe a me ka helu kope o ka gene target.Pono e noʻonoʻo ʻia ka nui hāpana e ke kūlana maoli.E hoʻoheheʻe i ka hāpana ma ka hoʻohui ʻana i ka wai nuclease-free a i ʻole TE Buffer, inā pono.

4. Paipai ʻia ka nui o nā laʻana like ʻole:

Ka Mana Mana

1. ʻIke hana: ʻike, kikoʻī a me ka hana hou ʻana o qPCR.

2. ʻAʻohe hana nuclease exogenous: ʻaʻohe exogenous endonuclease a me exonuclease pollution.

Nā memo o ka Huahana

1. Ke hoʻohana nei kēia huahana i kahi ʻano hou o ka DNA polymerase wela wela, hiki ke hoʻāla ʻia i nā minuke 1-5. No ka mea ua hoʻomaikaʻi ʻia kāna mea hoʻihoʻi, ʻoi aku ka maikaʻi no ka PCR quantitative fluorescence pālua a i ʻole me ka hoʻohana ʻana i ke ʻano probe.

2. Inā ha'aha'a loa ka waiwai Rn o ka ho'onui PCR a 'a'ole maopopo ka ho'onui 'ana, e ho'emi ana i ka nui o ka hāpana, e ho'onui i ka nui o ka hopena a i 'ole ka ho'ohehe'e mua 'ia o ka hāpana hiki ke ho'omaika'i i nā hopena.

3. ʻO ka hōʻiliʻili ʻana o ke koko, kohu, ka mimi, ka ʻāʻī ʻāʻī, a me nā mea ʻē aʻe e pono e hahai i nā koi koʻikoʻi o ka lāʻau lapaʻau, a hiki ke hoʻohana ʻia kahi laʻana hou e pale ai i ka degradation nucleic acid.

4. No ka loa'a 'ana o nā amplicons like 'ole i ka pono o ka ho'ohana 'ana i ka dUTP a me ka na'auao i ka UNG, pono e ho'onui 'ia nā reagents inā e emi ana ka 'ike i ka wā e ho'ohana ai i ka pūnaewele UNG.E ʻoluʻolu e kelepona mai iā mākou no ke kākoʻo ʻenehana inā pono.

5. No ka pale ʻana i ka hoʻonui ʻia ʻana o nā huahana PCR i lawe ʻia ma waena o nā hopena hoʻokahi, pono ka wahi hoʻokolohua i hoʻolaʻa ʻia a me ka pipette no ka hoʻonui ʻana.E hana me nā mīkina lima a hoʻololi pinepine a mai wehe i ka paipu hopena ma hope o ka hoʻonui ʻana o PCR.