QuickEasyᵀᴹ Wā Tūturu PCR Kit-Taqman
Whakaahuatanga kete:
◮Ko te punaha arotautanga PCR ahurei ka hanga 2×QuickEasyTMTino PCR Mix-Taqman he hototahi ake.
◮Ko te Hot-start Foregene HS Taq Polymerase he pai ake te kaha whakakaha, teitei ake te tairongo whakakaha, me te motuhake whakakaha ake.
◮2× TereTMReal PCR Mix -Ka whakamahi a Taqman i tetahi punaha ahurei i arotauhia e Foregene ki te whakapai ake i te maaramatanga me te motuhake o te rapunga rangahau-a-raupapa, ka taea te whakamahi mo te taatai me te whakatau i nga rereketanga tau kape me te whiwhi hua tika.
◮Ko tenei hua ka tae mai me te tae tohutoro o roto o te ROX, ka taea te whakamahi hei whakakore i te papamuri tohu me te hapa tohu i waenga i nga puna, he pai mo nga kaihoko ki te whakamahi i nga momo taputapu PCR ine.
Whakaahuatanga
Ko te 2× QuickEasyTMReal PCR Mix-Taqman i whakaratohia e te QuickEasyTMKo te kete PCR Waa Tuturu-Taqman kete he reanga hou o te punaha whakauru ariki i hangaia mo te tauhohenga whakakaha PCR Wā Tuturu ma te whakamahi i nga rangahau rewharewha motuhake.Ko tana matua o Foregene HS Taq DNA Polymerase i ahu mai i te whakarereketanga Antibody, i whakamahia i te taha o Foregene i te punaha PCR kua arotauhia ake, he teitei ake te motuhake me te kaha whakakaha.Ka whakatauritea ki te ranunga PCR noa, he kaha ake te kaha o te whakakaha me te iti o te reeti koretake.Ka taea te whakamahi i roto i te tauhohenga PCR ine rewharewha hei whakaiti i te whakakaha kore-motuhake me te whakapai ake i te tika o te PCR.Ka taea e ia te whakapai ake i te motuhake o nga hua me te tauhohenga.I taua wa ano, i whakawhiwhia a ROX hei waikano tohutoro o roto.
Ko te QuickEasyTMKo te kete PCR Kit-Taqman mo te wa tika he whanui te tono ki te nui o te tauira, a ka taea e ia te tiki i te anau paerewa pai i roto i nga rohe ine rereke, ka taea te ine me te kite i nga ira whainga me te pai o te tukurua me te tino maia.
Whakatakotoranga
200 Whakaritenga,500 Whakaritenga,1000 Whakaritenga,2000 Whakaritenga
Nga waahanga kete
| TereEasyTMWaa Tuturu PCR Kit-Taqman(20μlpūnaha) | ||||
| Kei roto kete | QP-01121 | QP-01122 | QP-01123 | QP-01124 |
| 200 Whakaritenga | 500 Whakaritenga | 1000 Whakaritenga | 2000 Whakaritenga | |
| 2×TereMāmāTM PCR tuturu Whakaranu-Takamana | 1 mL×2 | 1.7 mL×3 | 1.7 mL×6 | 1.7 mL×12 |
| 20×Waikano Tohutoro ROX | 200μL | 0.5 mL | 1 mL | 1 mL×2 |
| ddH-korekore DNA2O | 1.7 mL | 1.7 mL×2 | 10 mL | 20 mL |
| IFU | 1wahi | 1wahi | 1wahi | 1wahi |
Nga waahanga me nga painga
■ Ko te punaha arotautanga PCR ahurei hanga 2×QuickEasyTMTino PCR Mix-Taqman he hototahi ake.
■ Hot-start Foregene HS Taq Polymerase he pai ake te kaha whakakaha, teitei ake te tairongo whakakaha, me te tino whakakaha ake.
■ 2× TereEasyTM Real PCR Mix -Ka whakamahi a Taqman i tetahi punaha ahurei i arotauhia e Foregene ki te whakapai ake i te maaramatanga me te motuhake o te rapunga rangahau-a-raupapa, ka taea te whakamahi mo te taatai me te whakatau i nga rereketanga tau kape me te whiwhi hua tika.
■ Ka tae mai tenei hua me te tae tohutoro o roto o te ROX, ka taea te whakamahi ki te whakakore i te papamuri tohu me te hapa tohu i waenga i nga puna, he pai mo nga kaihoko ki te whakamahi i nga momo taputapu PCR ine.
Te tono kete
■ Korekorero PCR ine mo te tauira tātari ine;
■ Cte whakanui PCR tikanga;
■ Ka taea te whakamahi mo te rapu allele.
Rokiroki me te ora o te Papa
Me penapena te kete ki te -20°C i roto i te pouri;Mena ka whakamahia i nga wa maha, ka taea hoki te rongoa ki te 4 ° C mo te wa poto (whakamahia i roto i nga ra 10).
Tikanga Tuturu PCR maataapono hoahoa tuatahi
Primer Whakamua me te Primer Whakamuri
Mo PCR Wā Tuturu, he mea nui te hoahoa tuatahi.Ko nga kaupapa tuatahi e pa ana ki te motuhake me te kaha o te whakanui PCR, a ka taea te hoahoa ma te tohu ki nga kaupapa e whai ake nei:
Te roa o te tuatahi: 18-30bp.
Te ihirangi GC: 40-60%.
Uara Tm: Ka taea e te rorohiko hoahoa tuatahi, penei i te Primer 5, te uara Tm o te tuatahi.Ko nga uara Tm o nga timatanga o runga me te taha whakararo kia tata tonu.Ka taea hoki te whakamahi i te tauira tatauranga Tm: Tm = 4 °C (G + C) + 2 °C (A + T).I te wa e mahi ana i te PCR, ka tohua te pāmahana i raro iho i te uara Tm tuatahi o te 5 °C hei pāmahana whakaenee (ka taea e te pikinga o te pāmahana whakaenee te whakanui ake i te motuhake o te tauhohenga PCR).
Nga mea tuatahi me nga hua PCR:
Ko te roa o te hua whakakaha PCR tuatahi he pai ki te 100-150bp.
Me karohia nga tauira hoahoa i te waahanga hanganga tuarua o te tauira ka taea.
Me karo i te hanga 2 neke atu ranei nga turanga taapiri i waenga i nga pito 3′ o nga puna tuatahi o te awa ki raro.
Kare e taea e te turanga tauranga tuatahi 3′ te noho me te 3 taapiri G, C ranei.
Ko nga mea tuatahi kaore he hanganga taapiri, ki te kore ka hangaia he hanganga makawe, ka pa ki te whakanui PCR.
Me tohatoha te ATCG kia rite ki te raupapa tuatahi, me karohia te turanga 3′ hei T.
Tāpiritanga 1: Pūtau Direct RT-qPCR Kit mōkihi tāpiri wāhanga
1.Rongoa Raihana Pūtau
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| |||
| Nga waahanga kete (24-puna puna lysis / puna) | DRT-01011-A1 | DRT-01011-A2 | |
| 100 T | 500 T | ||
| WahiI | Kaipupuri CL | 20 ml | 100 ml |
| Foregene Protease Plus II | 400 μl | 1 ml × 2 | |
| Kaipupuri ST | 1 ml × 2 | 10 ml | |
| WahiII | Pūkōmuru DNA | 400 μl | 1 ml × 2 |
2.RT Mix
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| |
| Nga waahanga kete (20 μl pūnaha tauhohenga) | DRT-01011-B1 |
| 200 T | |
| 5× Direct RT Mix | 800 μl |
| RNase-Free ddH2O | 1.7 ml × 2 |
|
| ||
| Nga waahanga kete (20 μl pūnaha tauhohenga) | DRT-01011-C1 | DRT-01011-C2 |
| 200 T | 1000 T | |
| 2× Tikanga qPCR Mix-SYBR | 1 ml × 2 | 1.7 ml × 6 |
| 50× ROX Waikano Tohutoro | 40 μl | 200 μl |
| RNase-Free ddH2O | 1.7 ml | 10 ml |
