Yakananga RT-qPCR Kit III
Kit Description:
◮Yakananga RT-qPCR Kit, uchishandisa Foreasy Reverse Transcriptase uye Foreasy HS Taq DNA Polymerase inogadzirwa neForegene, ine yakasarudzika reaction buffer, inoita kuti kit iyi ive nekupikisa kwakasimba uye kuenderana, uye inogona kuyedza zvakananga kuita kushandisa Foregene Lysis system setemplate.Iyi kit inogona kunyorera COVID-19 nucleic acid yekuona kit uye mamwe pathogenic bhakitiriya nucleic acid yekuona kit yekuvandudza.
Tsanangudzo
Yakananga RT-qPCR Kit inopa yakarongedzerwa RT-qPCR buffer (ine Foreasy HS Taq DNA polymerase yakasanganiswa) uye yakanyanya kushanda.Foreasy Reverse Transcriptase(MMLV),zvichiita kuti zvive nyore kugadzirwa kwemashure-ekupedzisira kits uye kugadzirisa system, mukati mekusimbisa kuri nyore.
Yakananga RT-qPCR Kit, uchishandisa Foreasy Reverse Transcriptase uye Foreasy HS Taq DNA Polymerase inogadzirwa neForegene, ine yakasarudzika reaction buffer, inoita kuti kit iyi ive nekupikisa kwakasimba uye kuenderana, uye inogona kuyedza zvakananga kuita kushandisa Foregene Lysis system setemplate.Iyi kit inogona kunyorera COVID-19 nucleic acid yekuona kit uye mamwe pathogenic bhakitiriya nucleic acid yekuona kit yekuvandudza.
Iyi kit inogona zvakananga kuve chikamu chechigadzirwa cheIVD, uchishandisa zviri nyore uye nekukurumidza.Inongoda kuongororwa nyore, pasina kuvandudza zvakare.
Zvinotsanangurwa
500T, 50,000T
Kit zvikamu
| Kit zviri mukati (20 μL reaction system) | IM-05111-03 | IM-05112-03 |
| 500 T | 50,000 T | |
| Foreasy REverse Transcriptase(200 U/μL) | 50 μL×1 | 1 mL×5 |
| 2 × Yakananga qPCRMix-Taqman | 1 mL ×5 | 500 mL×1 |
| Instruction | 1 | 1 |
Matanho Okushanda
A: Gadzirira template uye reagent
1. Gadzirira yakagadzirirwa RNA template (Inokurudzirwa kushandisa Foregene Total RNA Isolation Kit series kit kuti ubvise uye uchenese RNA template) kana sampuli yekugadzira chigadzirwa (Inokurudzirwa kushandisa Foregene Lysis system), mapeji chaiwo (10 μM) uye zvimwe zvinoshandiswa zvinoshandiswa, chiridzwa.
2. Isa 2 × Direct RT-qPCR Buffer, RNase-Free ddH2O uye 20 × ROX Reference Dye (kana ichidiwa) mubhokisi nechando, zvichiita kuti izvi zvinyunguduke zvakasikwa, uye zvakasanganiswa zvinyoro.
B: Gadzirira yeRT-qPCR system
Tora hafu yevhoriyamu yekupindura buffer ye reaction system (semuenzaniso, kana huwandu hweiyo system iri 20 μL, inoda kuwana gumi μL 2 × Yakananga RT-qPCR Buffer.). Foryakaenzana huwandu hwe enzyme,tinokurudzira kutiM-MLV: 10-30 U/20 μL maitiro ekuita system,TaqDNA polymerase:1-2 U / 20 μL maitiro maitiroiyi izano redu, iwe unofanirwa kuyedza nekuigadzirisa), wedzera RNA template, chaiyo primers uye probes, uye wedzera RNase-Yemahara ddH2O kusvika ku20 μL.Iyo chaiyo gadziriro yeRT-qPCR reaction system inogona kuendeswa kune inotevera tafura 1.
Tafura 1: Gadzirira yeRT-qPCR system
| Chikamu | Volume | Final concentration |
| 2 × Yakananga RT-qPCR Buffer | 10 μL | 1× |
| Foreasy Reverse Transcriptase(MMLV) | 10-30 U | |
| Foreasy HS Taq DNA Polymerase | 1-2 U | |
| Pamberi Pekutanga (10 μM) | 0.8 μL | 50-900nM |
| Reverse Primer(10 μM) | 0.8 μL | 50-900nM |
| Probe (4 μM) | 1 μL | 200nM |
| Template (RNA kana Lysate) | X μL | |
| 20 × ROX Reference Dye | - | 1* |
| RNase-FreeddH2O | (6.4-X) μL | |
| Total Volume | 20 μL |
Cherechedza: Forward Primer uye Reverse Primer ndiwo maprimers ejini rinotangwa.Iyo sisitimu yeqPCR inogona kugadziridzwa zvinoenderana neanoshanda ekuyedza uye PCR modhi.Isu tinokurudzira 400nM yekusangana kwekupedzisira kweakawanda maprimers.Rongedza vhoriyamu yeakananga maprimers uye probes zvinoenderana neyakagadzirirwa kusungwa uye yakakurudzirwa yekupedzisira yekumisikidza, ndapota.
1 *: Sarudza yakakodzera yekupedzisira kusungirirwa kweROX Reference Dye zvinoenderana nehuwandu hwakasiyana hwePCR zviridzwa.Iyo yakakwana ROX Reference Dye kuunganidzwa kwezvakajairwa kuwanda PCR zviridzwa inoratidzwa mutafura inotevera:
| Quantitative PCR zviridzwa | Yekupedzisira kusangana kweROX Reference Dye |
| ABI PRISM 7000/7300/7700/7900HT/Nhanho yekutanga,etc. | 1 × (semuenzaniso, wedzera 1μl 20 × ROX Reference Dye mu 20 μL reaction system) |
| ABI 7500, 7500 Fast, Stratagene Mx3000P, Mx3005P uye Mx4000, nezvimwewo. | 0.5× (semuenzaniso, wedzera 0.5μl 20×ROX Reference Dye mu 20 μL reaction system) |
| Roche PCR chiridzwa, Bio-Rad PCR chiridzwa, Eppendorf yakawanda PCR chiridzwa, nezvimwe. | Pasina ROX Reference Dye |
Storage uye Sherufu upenyu
Iyo kit inofanira kuchengetwa pa -20 ± 5 ℃.Inofanirwa kuchengetwa mu -20 ℃ thermostatic firiji.Nguva yechokwadi ndeye 2 makore kana yakachengetwa mune yakakodzera mamiriro.
