Ib-kauj ruam Rapid Mycoplasma Detection Kit (Isothermal Amplification Method)

Cov khoom siv tuaj yeem tshawb pom yim yam ntawm mycoplasma hauv cell culture nruab nrab los ntawm isothermal amplification txheej txheem thiab pom cov xim hloov. Yog tsis muaj DNA rho tawm, 1 μL cell kab lis kev cai supernatant tau ncaj qha ntxiv rau cov tshuaj tiv thaiv thiab 30 min tau tshwm sim ntawm 65 degree. Cov xim ntawm cov qauv zoo hloov los ntawm daj mus rau xim liab, thiab tag nrho cov txheej txheem tsis xav tau PCR ntsuas lossis qhib lub hau rau electrophoresis, uas tuaj yeem tiv thaiv cov kab mob sib kis los ntawm aerosol. Piv nrog rau cov txheej txheem PCR ib txwm muaj, cov khoom no muaj qhov zoo ntawm kev ua haujlwm yooj yim, siab rhiab heev, thiab tuaj yeem tiv taus ntau yam PCR inhibitors hauv kab lis kev cai nruab nrab, yog li tsis muaj qhov tshwm sim tsis zoo, thiab cov txiaj ntsig tshawb pom tau zoo ib yam nrog qPCR.

Xa nrog cov dej khov ntub thiab khaws cia ntawm -20 degree ; siv tau mus txog 12 lub hlis.

a: Muaj cov tshuaj chromogenic.

Ua ntej pib (thov ua tib zoo nyeem)

Teem 65 degree dej da dej lossis hlau cua sov thaiv ua ntej.

1. Cov qauv npaj yuav raug kuaj:

ib. Adherent cells: Absorb tus supernatant ncaj qha. Nws tau pom zoo tias cov qauv yuav tsum tau coj mus rau thaum lub cell inoculation los yog kua hloov ntau tshaj 3 hnub thiab qhov confluence degree yog hais txog 90%. Lub sijhawm no, cov ntsiab lus ntawm mycoplasma hauv supernatant yog siab thiab tuaj yeem tshawb pom tau yooj yim.

b. Cov hlwb raug tshem tawm: Cov supernatant tau txais tom qab centrifugation ntawm 1, 000 rpm rau 5 min. Nws tau pom zoo tias cov qauv yuav tsum tau muab coj los kuaj tom qab cell inoculation lossis kua dej hloov ntau dua 3 hnub, thaum cov ntsiab lus ntawm mycoplasma hauv cell kab lis kev cai nruab nrab siab thiab tuaj yeem kuaj pom tau yooj yim.

2. Cov tshuaj tiv thaiv kab mob

a: Mycored Buffer yog siv upside down tom qab thawing.

b: Raws li tus naj npawb ntawm cov qauv, tom qab cov tshuaj tiv thaiv saum toj no tau npaj, sub-pack rau hauv cov tshuaj tiv thaiv raj, ntxiv 20 μL ntawm Paraffin Roj, thiab tom qab ntawd mus rau theem tom ntej.

3. Ntxiv cov qauv

ib. Kev tswj tsis zoo: Tsis muaj cov qauv ntxiv rau thawj lub raj tshuaj tiv thaiv raws li kev tswj tsis zoo.

b. Kev tswj qhov zoo: 1 μL Kev Tswj Xyuas Zoo tau muab ntxiv rau qhov kawg cov tshuaj tiv thaiv raj raws li kev tswj tau zoo.

c. Qauv yuav tsum tau kuaj: Ntxiv 1 μL cell kab lis kev cai supernatant mus rau tus so ntawm cov tshuaj tiv thaiv raj.

a: Thov txuas lub taub hau ntawm cov lus qhia pipette nyob rau hauv cov kua theem ntawm paraffin oill ntxiv cov qauv.

4. Kev mob tshwm sim

Hloov cov tshuaj tiv thaiv raj mus rau lub preheated 65 degree dej da dej los yog hlau da dej rau incubate 30min. Yog tias qhov kev loj hlob xim tsis pom tseeb, lub sijhawm tshuaj tiv thaiv tuaj yeem txuas ntxiv mus rau 40 min.

Tshem tawm lub raj tshuaj tiv thaiv tam sim tom qab qhov kawg ntawm cov tshuaj tiv thaiv, thiab yog tias cov tshuaj tiv thaiv cov tshuaj tiv thaiv tseem daj tom qab nws rov qab los rau hauv chav tsev kub, qhov tshwm sim yuav tsis zoo. Yog hais tias cov tshuaj tiv thaiv yog xim liab, qhov tshwm sim yog qhov zoo. Lub raj tshuaj tiv thaiv yuav tsum tsis txhob qhib, txwv tsis pub nws yuav yooj yim ua rau muaj kuab paug aerosol.

1. Tsis txhob qhib lub raj tshuaj tiv thaiv tom qab cov tshuaj tiv thaiv dhau lawm.

2. Nws tau pom zoo tias cov lus qhia pipette nrog cov khoom lim yuav tsum tau siv rau hauv cov txheej txheem ntawm kev npaj cov tshuaj tiv thaiv, thiab cov lus qhia pov tseg pipette thiab cov tshuaj tiv thaiv lub raj yuav tsum muab tso rau hauv lub hnab ntim tus kheej thiab kho raws sijhawm.

3. Thov siv nucleic acid scavenger (pom zoo G3020) los so lub rooj zaum xeem thiab pipette nquag.

Rau Kev Tshawb Fawb Siv Tsuas!

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