Multiplex Otu Nzọụkwụ RT-qPCR Premix
Nkọwa
Nọmba nwamba: HCR5141A
Multiplex One Nzọụkwụ RT-qPCR Premix bụ PCR ọnụ ọgụgụ multiplex dabere na RNA dị ka ndebiri.N'ime nnwale ahụ, a na-eme ntụgharị ntụgharị na ọnụọgụ PCR n'otu tube mmeghachi omume ahụ, na-eme ka ọrụ nnwale dị mfe ma belata ohere nke mmetọ.Enwere ike iji nhazi pụrụ iche nke ihe nchekwa na ngwakọta enzyme mee ihe n'ime usoro lyophilized otu nzọụkwụ.Ngwa ahụ na-eji Reverse Transcriptase na-eguzogide okpomọkụ maka nhazi nke ọma nke eriri cDNA mbụ site na iji hotstart Taq DNA Polymerase maka mmụba ọnụọgụ.Ọ nwere ihe nchekwa mmeghachi omume kachasị mma, ngwakọta enzymes wdg, yana ihe ndị na-egbochi nkwalite PCR na-abụghị nke akọwapụtara ma na-abawanye arụmọrụ nke ọtụtụ mmeghachi omume qPCR agbakwunyere, na-eme ka mmụba dị ukwuu nke fluorescence na-eme ka ọ dị mma ma na-ahụ na nrụpụta nrụpụta nke primers.
Ngwa
| Aha |
| 1. Lyo-Buffer |
| 2. Ngwakọta Lyo-Enzyme |
| 3. Lyo nchebe |
Ọnọdụ ụgbọ njemion
A: Lyo-buffer na nchebe: -25 ~ -15 ℃, shelf ndụ bụ 1 afọ.
B: Ngwakọta Lyo-enzyme, 2-8 ℃, ndụ nchekwa bụ ọnwa 6.
Ntuziaka maka ịrụ ọrụ
1. Reaction System (Were 25μL dị ka ihe atụ)
| Ngwa | Olu (μL) | Ntụkwasị obi ikpeazụ |
| Lyo-Buffer | 6 | 1* |
| Ngwakọta nke Lyo-Enzyme | 1 | - |
| Lyo-echebe | 8 | - |
| Ngwakọta mbụ (10μM) | 1 | 0.1-1 uM |
| Ngwakọta nyocha (10μM) | 0.5 | 0.05-0.5uM |
| Ụdị RNA | 5 | - |
| DEPC H2O | Ruo 25 | - |
2. Protocol ịgba ígwè kachasị mma
1) Ụkpụrụ ịgba ígwè ọkọlọtọ
| Ogbo mmeghachi omume | Okpomọkụ | Oge | okirikiri | |
| 1 | Ntugharị azụ azụ | 50°Ca | Nkeji 10 | 1 |
| 2 | denaturation nke mbụ | 95°C | 5min | 1 |
| 3 | Mmeghachi omume nkwalite | 95°C | Nkeji 15 | okirikiri 45 |
| 60°Cb | 30 sekọndc |
2) Protocol ịgba ígwè ngwa ngwa
|
| Ogbo mmeghachi omume | Okpomọkụ | Oge | okirikiri |
| 1 | Ntugharị azụ azụ | 50°Ca | Nkeji 2 | 1 |
| 2 | denaturation nke mbụ | 95°C | 2 sekọnd | 1 |
| 3 | Mmeghachi omume nkwalite | 95°C | 1 sk |
okirikiri 45 |
| 60°Cb | Nkeji 13c |
Mara:
a) tụgharịa ederede: Okpomọkụ nwere ike họrọ 42°C ma ọ bụ 50°C maka 10-15 nkeji.
b) Mmeghachi omume nkwalite: A na-emezi ọnọdụ okpomọkụ dị ka uru Tm nke primers emebere.
c)Fluorescence mgbaàmà nnweta: Biko tọọ usoro nnwale dịka ihe achọrọ si dịakwụkwọ ntuziaka.
Ozi nka na ụzụ/ nkọwapụta
| Mmalite na-ekpo ọkụ | Nmalite ọkụ arụrụ arụ |
| Ụzọ nchọpụta | Nchọpụta nchọpụta mbụ |
| PCR usoro | Otu nzọụkwụ RT-qPCR |
| Ụdị ihe atụ | RNA |
Ihe ndetu
1. Ngwaahịa a bụ maka nyocha naanị.
2. Biko yiri PPE dị mkpa, uwe ụlọ nyocha na uwe aka, iji hụ na ahụike na nchekwa gị!
