中文Ngwa ihe nchọpụta RT-PCR
Aha ngwaahịa
Ngwa ihe nchọpụta RT-PCR nke ahụ ọkụ ezì (Lyophilized)
Nha
48 ule / ngwa, 50 ule / ngwa
Ezubere iji
Ngwa a na-eji ezigbo fluorescent RT-PCR usoro iji chọpụta RNA nke nje virus Swine fever (CSFV) na ihe ọrịa anụ ahụ dị ka tonsils, lymph nodes na splin na ihe ọrịa mmiri mmiri dị ka ọgwụ mgbochi na ọbara ezi.Ọ dabara maka nchọpụta, nchoputa na nchọpụta ọrịa gbasara nje virus Swine fever.Ngwa ahụ bụ sistemu PCR dị njikere (Lyophilized), nke nwere DNA amplification enzyme, reverse transcriptase, ihe nchekwa mmeghachi omume, primers akọwapụtara na nyocha achọrọ maka nchọpụta PCR fluorescent.
Ọdịnaya ngwaahịa
| Ngwa | ngwugwu | nkọwapụta | Ngwakọta |
| Ngwakọta nke CSFV PCR | 1 × karama (Lyophilized ntụ ntụ) | 50Nnwale | dNTP, mgCl2, Ndị mbụ, Nyocha, Reverse Transcriptase, Taq DNA polymerase |
| 6 × 0.2ml 8 nke ọma warara tube(Lyophilized) | 48Nnwale | ||
| Njikwa ziri ezi | 1 * 0.2ml tube (lyophilized) | 10Nnwale | Plasmid nwere iberibe CSFV akọwapụtara |
| Ngwọta mgbaze | 1.5 ml nke Cryotube | 500ul | / |
| Njikwa adịghị mma | 1.5 ml nke Cryotube | 200ul | 0.9% NaCl |
Nchekwa & Ndụ shelf
(1) Enwere ike ibuga ngwa ahụ na okpomọkụ ụlọ.
(2) The shelf ndụ bụ 18 ọnwa na -20 ℃ na 12 ọnwa na 2 ℃ ~ 30 ℃.
(3) Hụ akara na kit maka ụbọchị mmepụta na ụbọchị njedebe.
(4) The lyophilized ntụ ntụ version reagent kwesịrị ịchekwa na -20 ℃ mgbe mgbasa na ugboro ugboro ifriizi-thaw kwesịrị ihe na-erughị 4 ugboro.
Ngwa ngwa
GENECHECKER UF-150, UF-300 ezigbo oge fluorescence PCR akụrụngwa.
Eserese ọrụ
a) Ụdị karama:
b) 8 ụdị tube nke ọma:
Pcr Amplification
AkwadoroỊtọ ntọala
| Nzọụkwụ | okirikiri | Okpomọkụ (℃) | Oge | Ọwa fluorescence |
| 1 | 1 | 50 | 8 min | |
| 2 | 1 | 95 | Nkeji 2 | |
| 3 | 40 | 95 | 5s | |
| 60 | 10s | Anakọta FAM fluorescence |
* Mara: A ga-anakọta akara ngosi nke ọwa fluorescence FAM na 60 ℃.
Nsonaazụ ule ntụgharị asụsụ
| Ọwa | Nkọwa nke nsonaazụ |
| Ọwa FAM | |
| Ct≤35 | CSFV dị mma |
| Undet | CSFV adịghị mma |
| 35 | Nsonaazụ enyo enyo, nwale* |
* Ọ bụrụ na nnwale nyocha nke chanel FAM nwere uru Ct ≤40 wee gosi usoro nkwalite ụdị “S” a na-ahụkarị, a na-atụgharị nsonaazụ ya ka ọ dị mma, ma ọ bụghị na ọ dị njọ.
