Otu nzọụkwụ ngwa ngwa RT-qPCR Probe Premix-UNG
Nọmba nwamba: HCR5143A
Otu nzọụkwụ RT-qPCR Probe Kit (MAka ngwa ngwa) bụ ngwa nyocha ngwa ngwa RT-qPCR dabere na otu plex ma ọ bụ multiplex quantitative PCR na-eji RNA dị ka ndebiri (dị ka nje RNA).Ngwaahịa a na-eji ọgbọ ọhụrụ Taq DNA Polymerase emezigharị mgbochi mmadụ yana otu nzọụkwụ raara onwe ya nye Reverse Transcriptase, yana ihe nchekwa kachasị maka nkwalite ngwa ngwa, nke nwere ọsọ nkwalite ngwa ngwa, arụmọrụ nkwalite dị elu na nkọwapụta.Ọ na-akwado nkwalite nguzozi na ma otu-plex na multiplex nke ihe nlele dị ala na nke dị elu na obere oge.
Ngwa
1.5 × RT-qPCR ihe nchekwa (U+)
2. Ngwakọta Enzyme (U+)
ndetu:
a.Ihe nchekwa 5×RT-qPCR (U+) gụnyere dNTP na mg2+;
b.Ngwakọta Enzyme (U+) gụnyere reverse transcriptase, Hot Start Taq DNA polymerase, RNase inhibitor na UDG;
c.Jiri ndụmọdụ na-enweghị RNase, EP tubes, wdg.
Tupu iji ya, gwakọta nke ọma ihe nchekwa 5×RT-qPCR (U+).Ọ bụrụ na enwere mmiri ozuzo ọ bụla ka agbazechara, chere ka ihe nchekwa ahụ laghachi n'ime ụlọ okpomọkụ, gwakọta ma gbazee, wee jiri ha mee ihe.
Ọnọdụ Nchekwa
A na-ebufe ngwaahịa ahụ na ice akọrọ ma nwee ike ịchekwa ya na -25 ~ -15 ℃ maka 1 afọ.
Ntuziaka
1. Mmeghachi omume Sistemu
| Ngwa | Olu (20 μl mmeghachi omume) |
| 2 ×RT-qPCR ihe nchekwa | 4μl |
| Ngwakọta Enzyme (U+) | 0.8 μl |
| Premagaba n'ihu | 0.1 ~ 1.0μM |
| Mgbanwe nke mbụ | 0.1 ~ 1.0μM |
| Ihe nyocha TaqMan | 0.05 ~ 0.25μM |
| Ụdị | X μL |
| Mmiri na-enweghị RNase | ihe ruru 25 μl |
Ihe edeturu: olu mmeghachi omume bụ 10-50μL.
2. Protocol ịgba ígwè (Standard)
| okirikiri nzọụkwụ | Okpomọkụ | Oge | okirikiri |
| Ntugharị ntụgharị | 55 ℃ | 10 min | 1 |
| Mbido Denaturation | 95 ℃ | 30 sk | 1 |
| Denaturation | 95 ℃ | 10 sk | 45 |
| Mwepu / Mgbatị | 60 ℃ | 30 sk |
| Protocol ịgba ígwè (Ọsọ) okirikiri nzọụkwụ |
Okpomọkụ |
Oge |
okirikiri |
| Ntugharị ntụgharị | 55 ℃ | 5 min | 1 |
| Mbido Denaturation | 95 ℃ | 5 s | 1 |
| Denaturation | 95 ℃ | 3 s | 43 |
| Mwepu / Mgbatị | 60 ℃ | 10 s |
ndetu:
a.Okpomọkụ ntụgharị ntụgharị dị n'etiti 50 ℃ ruo 60 ℃, ịba ụba okpomọkụ na-enyere aka ịkwalite ihe dị mgbagwoju anya yana ndebiri ọdịnaya CG dị elu;
b.Ọ dị mkpa ka emeziwanye ọnọdụ okpomọkụ kacha mma dabere na uru Tm nke primer, wee họrọ oge kacha nso maka mkpokọta mgbama fluorescence dabere na ngwa PCR Real Time.
Ihe ndetu
Biko yiri PPE dị mkpa, uwe ụlọ nyocha na uwe aka, iji hụ na ahụike na nchekwa gị!
