QuickEasyᵀᴹ Real Time PCR Kit-Taqman
Kufotokozera Kwa Zida:
◮Makina apadera okhathamiritsa a PCR amapanga 2×QuickEasyTMReal PCR Mix-Taqman imagwirizana kwambiri.
◮Hot-start Foregene HS Taq Polymerase ili ndi mphamvu zokulirapo, kukhudzika kwapamwamba, komanso kukwezedwa kwambiri.
◮2 × QuickEasyTMReal PCR Mix -Taqman amagwiritsa ntchito kachitidwe kapadera kokometsedwa ndi Foregene kuti apititse patsogolo kukhudzika ndi kutsimikizika kwa kafukufuku wotsatizana, womwe ungagwiritsidwe ntchito pozindikira ma genotyping ndikutsitsa manambala ndikupeza zotsatira zolondola.
◮Chogulitsachi chimabwera ndi utoto wamkati wa ROX, womwe ungagwiritsidwe ntchito kuchotsa maziko azizindikiro ndi zolakwika zazizindikiro pakati pa zitsime, zomwe ndizosavuta kuti makasitomala azigwiritsa ntchito mumitundu yosiyanasiyana ya zida za PCR.
Kufotokozera
2 × QuickEasyTMReal PCR Mix-Taqman yoperekedwa ndi QuickEasyTMReal Time PCR Kit-Taqman kit ndi m'badwo watsopano wosakaniza makina opangidwa kuti azitha kukulitsa mawu a Real Time PCR pogwiritsa ntchito ma probe a fulorosenti.Pakatikati pake Foregene HS Taq DNA Polymerase idakhazikitsidwa pakusintha kwa Antibody, komwe kumagwiritsidwa ntchito molumikizana ndi Foregene kukhathamiritsanso PCR system, imakhala ndi kutsimikizika kwapamwamba komanso kukulitsa bwino.Poyerekeza ndi kusakaniza wamba kwa PCR, ili ndi kuthekera kokulirapo komanso kutsika kosagwirizana.Itha kugwiritsidwa ntchito mu fluorescence kuchuluka kwa PCR kuti muchepetse kukulitsa kosakhazikika ndikuwongolera kulondola kwa PCR.Iwo akhoza kwambiri kusintha mankhwala mwachindunji ndi anachita tilinazo.Nthawi yomweyo, ROX idaperekedwa ngati utoto wamkati.
The QuickEasyTMReal Time PCR Kit-Taqman kit ili ndi mawonekedwe otakata ku kuchuluka kwa template, ndipo imatha kupeza njira yabwino yokhotakhota m'magawo osiyanasiyana ochulukira, ndipo imatha kuwerengetsa molondola ndikuzindikira majini omwe mukufuna ndikubwereza bwino komanso chidaliro chachikulu.
Zofotokozera
200 zokonzekera,500 Kukonzekera,1000 Zokonzekera,2000 Preps
Zida zamagulu
| QuickEasyTMReal Time PCR Kit-Taqman(20μldongosolo) | ||||
| Zamkatimu | QP-01121 | QP-01122 | QP-01123 | QP-01124 |
| 200 Preps | 500 Preps | 1000 Preps | 2000 Preps | |
| 2 × paMwamsangaZosavutaTM PCR yeniyeni Sakanizani- Taqman | 1 ml×2 | 1.7 mL×3 | 1.7 mL×6 | 1.7 mL×12 |
| 20×ROX Reference Dye | 200μL | 0.5 ml | 1 ml | 1 ml×2 |
| DNase-Free ddH2O | 1.7 mL | 1.7 mL×2 | 10 ml pa | 20 ml |
| IFU | 1chidutswa | 1chidutswa | 1chidutswa | 1chidutswa |
Mbali & ubwino
■ Makina apadera a PCR okhathamiritsa amapanga 2×QuickEasyTMReal PCR Mix-Taqman imagwirizana kwambiri.
■ Hot-start Foregene HS Taq Polymerase ili ndi kukweza kwapamwamba kwambiri, kukhudzika kwapamwamba, komanso kamvekedwe kake kapamwamba.
■ 2× QuickEasyTM Real PCR Mix -Taqman amagwiritsa ntchito kachitidwe kapadera kokometsedwa ndi Foregene kuti apititse patsogolo kukhudzika ndi kutsimikizika kwa kafukufuku wotsatizana, womwe ungagwiritsidwe ntchito pozindikira ma genotyping ndikutsitsa manambala ndikupeza zotsatira zolondola.
■ Chogulitsachi chimabwera ndi utoto wamkati wa ROX, womwe ungagwiritsidwe ntchito kuthetsa maziko azizindikiro ndi zolakwika za chizindikiro pakati pa zitsime, zomwe zimakhala zosavuta kuti makasitomala azigwiritsa ntchito mumitundu yosiyanasiyana ya zida za PCR.
Kit application
■ Fluorescence quantitative PCR yowunikira kuchuluka kwa template;
■ Ckukulitsa kwa PCR;
■ Itha kugwiritsidwa ntchito pozindikira ma allele.
Kusungirako ndi moyo wa alumali
Chidacho chiyenera kusungidwa pa -20 ° C mumdima;ngati imagwiritsidwa ntchito pafupipafupi, imathanso kusungidwa pa 4 ° C kwa nthawi yochepa (kugwiritsa ntchito mkati mwa masiku 10).
Real Time PCR primer design mfundo
Forward Primer ndi Reverse Primer
Kwa Real Time PCR, mapangidwe oyambira ndi ofunika kwambiri.Zoyambira zimayenderana ndi kutsimikizika komanso kuthekera kwa kukulitsa kwa PCR, ndipo zitha kupangidwa motengera mfundo izi:
Kutalika koyambirira: 18-30bp.
Zolemba za GC: 40-60%.
Mtengo wa Tm: Mapulogalamu opangira ma primer, monga Primer 5, amatha kupereka mtengo wa Tm woyambira.Makhalidwe a Tm oyambira kumtunda ndi kumtunda ayenera kukhala pafupi momwe angathere.Njira yowerengera ya Tm ingagwiritsidwenso ntchito: Tm = 4 °C (G + C) + 2 °C (A + T).Pochita PCR, kutentha pansi pa mtengo woyambira wa Tm wa 5 ° C nthawi zambiri kumasankhidwa ngati kutentha kwapang'onopang'ono (kuwonjezeka kofananirako kwa kutentha kwapang'onopang'ono kumatha kukulitsa momwe PCR imachitira).
Primers ndi zinthu za PCR:
Design primer PCR amplification product kutalika makamaka 100-150bp.
Zoyambira zojambula m'dera lachiwiri la template ziyenera kupewedwa momwe zingathere.
Pewani kupanga zoyambira ziwiri kapena kupitilira apo pakati pa 3′ malekezero oyambira kumtunda ndi kumunsi kwa mtsinje.
Primer 3′ terminal base sangakhalepo ndi 3 zowonjezera zotsatizana za G kapena C.
Zoyambira zokha sizingakhale ndi zida zowonjezera, apo ayi mawonekedwe atsitsi adzapangidwa, zomwe zimakhudza kukulitsa kwa PCR.
ATCG iyenera kugawidwa mofanana momwe mungathere poyambira, ndipo 3′ terminal base iyenera kupewedwa ngati T.
Zowonjezera 1: Cell Direct RT-qPCR Kit chigawo chowonjezera paketi
1.Cell Lysis Solution
|
| |||
| Zida zamagulu (24-chitsime lysis dongosolo / bwino) | Chithunzi cha DRT-01011-A1 | Chithunzi cha DRT-01011-A2 | |
| 100 T | 500 T | ||
| GawoIne | Mtengo CL | 20 ml | 100 ml |
| Foregene Protease Plus II | 400 μl | 1 ml × 2 | |
| Mtengo wapatali wa magawo ST | 1 ml × 2 | 10 ml | |
| GawoII | DNA Eraser | 400 μl | 1 ml × 2 |
2.RT Mix
|
| |
| Zida zamagulu (20 μl reaction system) | Chithunzi cha DRT-01011-B1 |
| 200 T | |
| 5 × Direct RT Mix | 800 ml |
| RNase-Free ddH2O | 1.7ml × 2 |
|
| ||
| Zida zamagulu (20 μl reaction system) | DRT-01011-C1 | DRT-01011-C2 |
| 200 T | 1000 T | |
| 2 × Mwachindunji qPCR Mix-SYBR | 1 ml × 2 | 1.7ml × 6 |
| 50 × ROX Reference Dye | 40 ml | 200 μl |
| RNase-Free ddH2O | 1.7 ml | 10 ml |
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