Direct RT-qPCR Kit II
ʻO ka wehewehe ʻana o ka pahu:
◮Direct RT-qPCR Kit, me ka hoʻohana ʻana i ka Foreasy Reverse Transcriptase a me ka Foreasy HS Taq DNA Polymerase e hoʻomohala ʻia e Foregene, me ka pale kūʻokoʻa kūʻokoʻa, hana i kēia kit me ke kūpaʻa ikaika a me ka hoʻohālikelike ʻana, a hiki iā ia ke hoʻāʻo pololei i ka hoʻohana ʻana i ka ʻōnaehana Foregene Lysis e like me ka laʻana.Hiki ke hoʻohana ʻia kēia pahu no ka pahu ʻike ʻike nucleic acid COVID-19 a me nā mea ʻē aʻe pathogenic bacteria nucleic acid detection kit development.
wehewehe
Hāʻawi ʻo Direct RT-qPCR Kit i hoʻopaʻa ʻokoʻa ʻia ʻo RT-qPCR buffer, Foreasy HS Taq DNA Polymerase kiʻekiʻe.a me Foreasy Reverse Transcriptase (MMLV),maʻalahi no ka hana ʻana i nā pahu hope a me ka hoʻoponopono ʻōnaehana, i loko o ka hōʻoia maʻalahi.
Direct RT-qPCR Kit, me ka hoʻohana ʻana i ka Foreasy Reverse Transcriptase a me ka Foreasy HS Taq DNA Polymerase e hoʻomohala ʻia e Foregene, me ka pale kūʻokoʻa kūʻokoʻa, hana i kēia kit me ke kūpaʻa ikaika a me ka hoʻohālikelike ʻana, a hiki iā ia ke hoʻāʻo pololei i ka hoʻohana ʻana i ka ʻōnaehana Foregene Lysis e like me ka laʻana.Hiki ke hoʻohana ʻia kēia pahu no ka pahu ʻike ʻike nucleic acid COVID-19 a me nā mea ʻē aʻe pathogenic bacteria nucleic acid detection kit development.
Hiki i kēia pahu ke lilo pololei i mea o ka huahana IVD, me ka maʻalahi a me ka wikiwiki.Pono wale ka hōʻoia maʻalahi, me ka hoʻomohala hou ʻole.
Nā kikoʻī
500T, 50,000T
Nā ʻāpana pahu
| Kiko maʻiʻo (20 μL ʻōnaehana pane) | IM-05111-02 | IM-05112-02 |
| 500 T | 50,000 T | |
| ʻO 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× pololei RT-qPCRHoʻopaʻa | 1 mL ×5 | 500 mL ×1 |
| Aʻo | 1 | 1 |
Nā ʻanuʻu hana
A: E hoʻomākaukau i ka template a me ka reagent
1. E hoʻomākaukau i ka laʻana RNA i hoʻomākaukau ʻia (Aia ʻia e hoʻohana i ka Foregene Total RNA Isolation Kit series kit e unuhi a hoʻomaʻemaʻe i ka RNA template) a i ʻole nā huahana hoʻoheheʻe ʻia (Ua paipai ʻia e hoʻohana i ka ʻōnaehana Foregene Lysis), nā primers kikoʻī (10 μM) a me nā mea hoʻohana e pili ana.
2. E kau i ka 2× Direct RT-qPCR Buffer, RNase-Free ddH2O a me 20× ROX Reference Dye (inā pono) i loko o ka pahu me ka hau, e hoohehee maoli ai keia mau mea, a hui malie.
B: E hoʻomākaukau i ka ʻōnaehana RT-qPCR
E kiʻi i ka hapalua o ka leo o ka pānaʻi ʻana o ka ʻōnaehana pane (no ka laʻana, inā he 20 μL ka nui o ka ʻōnaehana, pono e loaʻa iā 10 μL 2 × Direct RT-qPCR Buffer.). No ka meaka nui o ka enzyme, manaʻo mākou i kēlāM-MLV:10-30 U/20 μL ʻōnaehana pane,TaqDNA polymerase：1-2 U/20 μL ʻōnaehana paneʻO kēia kā mākou manaʻo, pono ʻoe e hoʻāʻo a hoʻoponopono), e hoʻohui i ka laʻana RNA, nā kumu mua a me nā ʻimi, a hoʻohui i ka RNase-Free ddH2ʻO ka 20 μL.Hiki ke kuhikuhi ʻia ka hoʻomākaukau kikoʻī o ka ʻōnaehana pane RT-qPCR i ka papa 1 aʻe.
Papa 1 : E hoʻomākaukau i ka ʻōnaehana RT-qPCR
| ʻāpana | Volume | Hoʻoholo hope |
| 2× Kuhi pololei RT-qPCR Buffer | 10 μL | 1× |
| Foreasy Reverse Transcriptase(MMLV) | 10-30 U | |
| Foreasy HS Taq DNA Polymerase | 1-2 U | |
| Kuhi mua (10 μM) | 0.8 μL | 50-900nM |
| Pumi Hoʻohuli (10 μM) | 0.8 μL | 50-900nM |
| ʻImi (4 μM) | 1 μL | 200nM |
| Palapala (RNA a i ʻole Lysate) | X μL | |
| 20× ROX Peʻa Waikane | - | 1* |
| RNase-FreeddH2O | (6.4-X) μL | |
| Huina Volume | 20 μL |
'Ōlelo Aʻo: Forward Primer a me Reverse Primer he mau kumu mua ia o ka gene target.Hiki ke hoʻololi ʻia ka ʻōnaehana o ka qPCR e like me nā hiʻohiʻona hoʻokolohua kūpono a me PCR.Manaʻo mākou iā 400nM no ka ʻike hope loa o ka hapa nui o nā primers.E hoʻololi i ka leo o nā primers kikoʻī a me nā ʻimi e like me ka ʻike i hoʻomākaukau ʻia a me ka ʻike hope i manaʻo ʻia, e ʻoluʻolu.
1*: E koho i ka noʻonoʻo hope kūpono o ROX Reference Dye e like me nā mea hana PCR quantitative like ʻole.Hōʻike ʻia ka manaʻo ROX Reference Dye maikaʻi loa o nā mea kani PCR quantitative maʻamau i ka papa ma lalo:
| Nā mea hana PCR nui | ʻO ka manaʻo hope loa o ROX Reference Dye |
| ABI PRISM 7000/7300/7700/7900HT/Step One,etc. | 1× (no ka laʻana, hoʻohui i 1μl 20×ROX Reference Dye ma 20 μL ʻōnaehana pane) |
| ABI 7500, 7500 Fast, Stratagene Mx3000P, Mx3005P a me Mx4000, etc. | 0.5× (no ka laʻana, hoʻohui i ka 0.5μl 20×ROX Reference Dye ma 20 μL reaction system) |
| ʻO Roche PCR mea kani, Bio-Rad PCR mea kani, Eppendorf quantitative PCR mea kani, etc. | Me ka ʻole ROX Reference Dye |
Ka mālama ʻana a me ka mālama ʻana
Pono e mālama ʻia ka pahu ma -20 ± 5 ℃.Pono e mālama koke ʻia i loko o ka friji thermostatic -20 ℃.ʻO ka manawa kūpono he 2 mau makahiki inā mālama ʻia i kahi kūlana kūpono.
