中文Ihe nchọpụta PCR nke Listeria monocytogenes
Aha ngwaahịa
Ihe nchọpụta PCR nke Listeria monocytogenes
Nha
48 ule / ngwa, 50 ule / ngwa
Ezubere iji
Listeria monocytogenes bụ microbacterium nwere gram nke nwere ike itolite n'etiti 4 ℃ na 45 ℃.Ọ bụ otu n'ime ihe ndị na-akpata ọrịa na-eyi ahụike mmadụ egwu na nri friji.Isi ngosipụta nke ọrịa bụ septicemia, meningitis na mononucleosis.Ngwa a dabara adaba maka nchọpụta qualitative nke Listeria monocytogenes na nri, ihe nlele mmiri, feces, vomit, mmiri mmiri na-eme ka nje bacteria na ihe atụ ndị ọzọ na-eji ụkpụrụ nke ezigbo oge fluorescence PCR. The kit bụ PCR-PRDY SYSTEM (Lyophilized) , nke nwere DNA amplification enzyme, reverse transcriptase, nzaghachi mmeghachi omume, kpọmkwem primers na nyocha achọrọ maka nchọpụta PCR fluorescent.
Nchekwa & Ndụ nchekwa
(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.
Ọdịnaya ngwaahịa
| Ngwa | ngwugwu | nkọwapụta | Ngwakọta |
| Listeria monocytogenes PCR Mix | 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 mpempe akwụkwọ Listeria monocytogenes |
| Ngwọta mgbaze | 1.5 ml nke Cryotube | 500ul | / |
| Njikwa adịghị mma | 1.5 ml nke Cryotube | 200ul | 0.9% NaCl |
Ngwa ngwa
GENECHECKER UF-150, UF-300 ezigbo oge fluorescence PCR akụrụngwa.
Eserese ọrụ
a)
b)
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 | Listeria monocytogenes dị mma |
| Undet | Listeria monocytogenes 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.
