Cov Khoom Siv Sib Nqus Rau Kab Mob DNA/RNA HYD521
Cov Khoom Siv Magnetic Universal Viral DNA/RNA Kit no yog siv rau kev rho cov kua qaub nucleic ntawm cov ntshav tag nrho lossis cov kua dej hauv lub cev uas tsis muaj cell (xws li cov serum, plasma, cerebrospinal fluid, nasal/pharyngeal swabs, alveolar lavage fluid, thiab lwm yam). Txoj kev no siv cov thev naus laus zis magnetic bead purification, thiab tsis tas yuav muaj tshuaj lom phenol chloroform extraction thaum lub sijhawm rho tawm. Nws muaj kev nyab xeeb, tsis muaj tshuaj lom thiab ceev ceev. Cov khoom tau txais tuaj yeem siv ncaj qha rau hauv PCR, qPCR, second-generation sequencing thiab lwm yam kev sim. Thaum siv nrog cov cuab yeej rho tawm tsis siv neeg magnetic bead, kev rho tawm cov kua qaub nucleic muaj zog tuaj yeem ua tiav.
Cov ntaub ntawv khoom
| Cov Cheebtsam | 48T |
| Kev daws teeb meem lysis thiab kev sib khi | 33 ml |
| Kev ncua ntawm cov hlaws dai sib nqus | 1.1 mL |
| Ntxuav lub tshuab ntxhua khaub ncaws A | 40 mL |
| Cov tshuaj ntxuav B | 80 ml |
| Kev daws teeb meem | 10 mL |
| Proteinase K | 1.1 mL |
Cov Cai Khaws Cia
Proteinase K yuav tsum khaws cia rau ntawm 2 ~ 8 ℃ thiab lwm cov khoom xyaw yuav tsum khaws cia rau ntawm chav sov. Lub sijhawm khaws cia yog 18 lub hlis.
Qauv Piav Qhia
1. Cov qauv kuaj siv tau: cov ntshav tag nrho, cov serum, plasma, cov kua hauv lub hlwb, cov ntaub so ntswg/pharyngeal, cov kua ntxuav alveolar thiab lwm yam qauv kuaj.
2. Kev khaws cia thiab thauj cov qauv kuaj: Cov qauv kuaj siv tau rau kev kuaj tam sim ntawd lossis khaws cia rau ntawm -70 ℃ lossis qis dua rau kev kuaj. Lub sijhawm khaws cia yog 6 lub hlis. Yuav tsum tsis txhob khov thiab yaj dua. Cov qauv kuaj raug thauj los ntawm cov saw txias.
3. Cov kev cai rau kev khov thiab yaj: Khov thiab yaj sai, tsis txhob khov thiab yaj dua.
Kev Ua Haujlwm
Ua ntej qhov kev sim, xyuas seb cov tshuaj puas muaj nag thiab seb cov hlaws hlau nplaum puas tuaj yeem rov ua dua.
1. Hloov 200~300 μL ntawm cov qauv mus rau hauv lub raj centrifuge 1.5 mL, ntxiv 600 μL ntawm Lysis thiab Binding Solution, 20 μL ntawm Proteinase K, thiab 20 μL ntawm cov hlau nplaum sib nqus ua ke, thiab vortex ntawm qhov ceev ceev rau 30 vib nas this. Incubate ntawm 50 ℃ rau 5 ~ 7 feeb. Yog tias lub thermostat tsis muaj oscillation function, vortex 3 zaug thaum lub sijhawm incubation, txhua zaus rau 15 vib nas this.
2. Muab tso rau hauv lub thawv sib nqus 1.5 mL rau kev sib cais sib nqus kom txog thaum cov tshuaj ntshiab thiab pob tshab, thiab pov tseg cov tshuaj.
3. Ntxiv 700 μL ntawm cov tshuaj ntxuav A, vortex rau 10 vib nas this kom tawg cov hlaws sib nqus, thiab tom qab ntawd hloov mus rau lub rooj sib nqus rau kev sib cais sib nqus kom txog thaum cov tshuaj ntshiab, thiab pov tseg cov tshuaj.
4. Ntxiv 700 μL ntawm cov tshuaj ntxuav B, vortex rau 10 s kom tawg cov hlaws sib nqus, hloov mus rau lub rooj sib nqus rau kev sib cais sib nqus kom txog thaum cov tshuaj ntshiab, thiab pov tseg cov tshuaj.
5. Ntxiv 700 μL ntawm cov tshuaj ntxuav B dua, vortex rau 10 s kom tawg cov hlaws sib nqus, hloov mus rau lub rooj sib nqus rau kev sib cais sib nqus kom txog thaum cov tshuaj ntshiab, thiab pov tseg cov tshuaj.
6. Siv lub tshuab centrifuge luv luv los sau cov tee dej ntawm phab ntsa ntawm lub raj, muab tso rau ntawm lub rooj sib nqus kom pom tseeb, thiab nqus cov kua seem. Qhuav rau 3-5 feeb.
Lus Cim: Cov khoom seem ethanol yuav tiv thaiv cov tshuaj enzyme tom qab ntawd, yog li xyuas kom meej tias ethanol ua pa tag thaum ziab. Tsis txhob ziab ntev dhau kom tsis txhob cuam tshuam rau cov txiaj ntsig elution tom qab.
7. Ntxiv 40~100 μL ntawm Elution solution, vortex ntawm qhov ceev ceev rau 2~3 feeb kom tawg cov hlaws magnetic. Incubate ntawm 60℃ rau 5 feeb, tom qab ntawd vortex ntawm qhov ceev ceev rau 60 vib nas this.
8. Siv lub tshuab centrifuge tam sim ntawd los sau cov kua los ntawm lub hau raj thiab hloov mus rau lub khib hlau nplaum kom txog thaum cov hlaws hlau nplaum raug nqus tag. Tom qab ntawd ua tib zoo hloov cov kua mus rau lub raj centrifuge tshiab kom tau txais cov kua nucleic acid.
9. Cov kua qaub nucleic acid tuaj yeem khaws cia rau -20 ° C rau kev khaws cia luv luv thiab -80 ° C rau kev khaws cia ntev.
Cov Lus Cim
1. Cov tshuaj buffers sib txawv hauv cov khoom siv no muaj cov ntsev guanidine. Rau koj txoj kev nyab xeeb thiab kev noj qab haus huv, thov hnav lub tsho tiv no kuaj mob thiab hnab looj tes pov tseg. Thiab tuav raws li cov qauv kev nyab xeeb. Tsis txhob cia cov tshuaj buffer kov rau ntawm daim tawv nqaij, qhov muag thiab cov ntaub so ntswg. Yog tias nws tshwm sim, thov ntxuav tam sim ntawd nrog dej ntau thiab nrhiav kev kho mob.
2. Yog tias cov tshuaj no poob rau hauv av, yuav tsum tau muab tso rau hauv dej 30 ℃ kom txog thaum cov dej ntawd yaj tag ua ntej siv.
3. Yog tias cov hlau nplaum uas raug tshem tawm khov lawm, tsis txhob siv nws.
4. Cov buffers sib txawv hauv cov khoom siv no muaj cov ntsev guanidine. Tsis txhob siv cov tshuaj tua kab mob oxidizing xws li sodium hypochlorite los kho lawv, txwv tsis pub cov pa phem yuav raug tso tawm thiab lawv yuav tsum tau kho raws li cov khib nyiab kho mob.
5. Tej zaum yuav muaj cov hlaws hlau nplaum seem thaum lub sijhawm elution. Sim tsis txhob nqus cov hlaws hlau nplaum thaum nqus cov qauv.