Ngwa ihe nchọpụta RT-PCR

Nkọwa dị mkpirikpi:

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.


Nkọwa ngwaahịa

Mkpado ngwaahịa

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:

1

b) 8 ụdị tube nke ọma:

2

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.


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