Tika RT-qPCR Kit II
Whakaahuatanga kete:
◮Tika RT-qPCR Kit, ma te whakamahi i te Foreasy Reverse Transcriptase me te Foreasy HS Taq DNA Polymerase e whakawhanake ana e Foregene, me te aukati tauhohenga ahurei, ka hangaia tenei kete ki te kaha te aukati me te hototahi, a ka taea e ia te whakamatautau tika te whakamahi i te punaha Foregene Lysis hei tauira.Ka taea e tenei kete te tono mo te kete kimi waikawa nucleic COVID-19 me etahi atu huakita pathogenic ki te whanaketanga kete kitenga waikawa karihi.
Whakaahuatanga
Ko te Kete RT-qPCR tika e whakarato ana i te parepare RT-qPCR kua taia motuhake, tino pai te Foreasy HS Taq DNA Polymeraseme Foreasy Reverse Transcriptase(MMLV),kia watea mo te hanga kete o muri-mutunga me te whakatikatika i nga punaha, i roto i te manatoko ngawari.
Tika RT-qPCR Kit, ma te whakamahi i te Foreasy Reverse Transcriptase me te Foreasy HS Taq DNA Polymerase e whakawhanake ana e Foregene, me te aukati tauhohenga ahurei, ka hangaia tenei kete ki te kaha te aukati me te hototahi, a ka taea e ia te whakamatautau tika te whakamahi i te punaha Foregene Lysis hei tauira.Ka taea e tenei kete te tono mo te kete kimi waikawa nucleic COVID-19 me etahi atu huakita pathogenic ki te whanaketanga kete kitenga waikawa karihi.
Ka taea e tenei kete te waahanga o te hua IVD, te whakamahi ngawari me te tere.Me ngawari noa te manatoko, me te kore e whanake ano.
Whakatakotoranga
500T, 50,000T
Nga waahanga kete
| Kei roto kete (20 μL pūnaha tauhohenga) | IM-05111-02 | IM-05112-02 |
| 500 T | 50,000 T | |
| Foreasy Reverse Transcriptase (200 U/μL) | 50 μL×1 | 1 mL×5 |
| Foreasy HS Taq DNA Polymerase (5 U/μL) | 100 μL×1 | 1 mL×10 |
| 2× Tika RT-qPCRKaipupuri | 1 mL ×5 | 500 mL ×1 |
| Tohutohu | 1 | 1 |
Nga Mahi Mahi
A: Whakaritea te tauira me te reagent
1. Whakapaia te tauira RNA kua rite (E taunaki ana kia whakamahia te kete raupapa Kete Isolation RNA Tapeke o Foregene ki te tango me te pure i te tauira RNA) he tauira hua kapiti ranei (E taunaki ana kia whakamahi i te punaha Foregene Lysis), nga mea tuatahi (10 μM) me etahi atu taonga e tika ana, taputapu.
2. Kuhua te Waikano Tohutoro 2× Direct RT-qPCR, RNase-Free ddH2O me te 20× ROX Reference Dye(mehemea e hiahiatia ana) ki roto i te pouaka me te hukapapa, kia rewa noa enei, ka uru ata.
B: Whakaritehia te punaha RT-qPCR
Tikina te haurua o te rōrahi pūrei tauhohenga o te pūnaha tauhohenga (hei tauira, ki te 20 μL te rōrahi o te pūnaha, me whiwhi 10 μL 2× Direct RT-qPCR Buffer). Mote nui o te whākōkī, e whakaaro ana matouM-MLV:10-30 U/20 μL pūnaha tauhohenga,TaqDNA polymerase：1-2 U/20 μL pūnaha tauhohengaKo ta matou whakaaro tenei, me whakamatau koe me te whakatika), taapirihia te tauira RNA, nga tohu tuatahi me nga rangahau, me te taapiri i te RNase-Free ddH2O ki te 20 μL.Ko te whakaritenga motuhake o te punaha urupare RT-qPCR ka taea te tuku ki te ripanga 1 e whai ake nei.
Ripanga 1 : Whakaritehia te punaha RT-qPCR
| Waehanga | Rōrahi | Te kukū whakamutunga |
| 2× Tika RT-qPCR Pūreirei | 10 μL | 1× |
| Foreasy Whakamuri Transcriptase(MMLV) | 10-30 U | |
| Foreasy HS Taq DNA Polymerase | 1-2 U | |
| Pūmua Whakamua(10 μM) | 0.8 μL | 50-900nM |
| Pūmatua Whakamuri(10 μM) | 0.8 μL | 50-900nM |
| Probe(4 μM) | 1 μL | 200nM |
| Tātauira(RNA, Lysate ranei) | X μL | |
| 20× ROX Waikano Tohutoro | - | 1* |
| RNase-FreeddH2O | (6.4-X) μL | |
| Rōrahi Tapeke | 20 μL |
Tuhipoka: Ko te Kuputuhi Whakamua me te Panui Whakamuri he tohu tuatahi o te ira e whai ana.Ko te punaha o te qPCR ka taea te whakatika kia rite ki nga tauira whakamatautau me te PCR.Ka tūtohu matou ki te 400nM mo te kukū whakamutunga o te nuinga o nga tuatahi.Whakaritea te rōrahi o ngā pūmua me ngā rangahau kia rite ki te kukū kua whakaritea me te kukū whakamutunga e tūtohu ana, tēnā.
1*: Tīpakohia te kukū whakamutunga o te Waikano Tohutoro ROX kia rite ki nga taputapu PCR ine.Ko te tino pai o te Rox Reference Dye concentration o nga taputapu PCR ine noa e whakaatuhia ana i te ripanga e whai ake nei:
| Nga taputapu PCR ine | Te kukū whakamutunga o te Waikano Tohutoro ROX |
| ABI PRISM 7000/7300/7700/7900HT/Taahiraa Tuatahi,etc. | 1× (hei tauira, tāpirihia te Waikano Tohutoro 1μl 20×ROX ki te pūnaha tauhohenga 20 μL) |
| ABI 7500, 7500 Tere, Stratagene Mx3000P, Mx3005P me Mx4000, etc. | 0.5× (hei tauira, tāpirihia te Waikano Tohutoro 0.5μl 20×ROX ki te pūnaha tauhohenga 20 μL) |
| Roche PCR taputapu, Bio-Rad PCR taputapu, Eppendorf quantitative PCR taputapu, etc. | Kore he Waikano Tohutoro ROX |
Rokiroki me te ora o te Papa
Me pupuri te kete ki te -20 ± 5 ℃.Me pupuri tonu i roto i te -20 ℃ pouaka whakamatao.Ko te wa mana he 2 tau mena ka penapena i roto i nga ahuatanga pai.
