PCR vs Real-time PCR
PCR lossis Polymerase cov tshuaj tiv thaiv kab mob yog qhov kev tshawb pom nyob rau hauv niaj hnub molecular biology, uas yog thawj zaug tsim los ntawm chemist Kary Mullis hauv xyoo 1983. Nws tso cai rau ib ntu hauv cov DNA nyuaj kom nthuav dav rau kev tshuaj xyuas. Lub tswv yim tseem ceeb ntawm PCR yog tias ob lub primers, uas yog ntxiv rau qhov sib txawv ntawm DNA ib ntus, yog taw qhia rau ib leeg; cov primers tsim cov strands ntxiv, txhua tus muaj lwm cov primer. Yog li, qhov tshwm sim yog qhov ntau ntawm cov kab ke sib txuas rau DNA uas nyob nruab nrab ntawm ob lub primers. DNA polymerase enzyme yog siv los txuas ntxiv cov primers hauv PCR. DNA polymerase yog ib tug thermostable enzyme, thiab nws muaj peev xwm ciaj sia nyob rau hauv siab kub (94 mus rau 95 ° C) siv rau denaturation ntawm template DNA.
PCR suav nrog peb kauj ruam, uas yog, rov ua qhov rov ua dua tshiab, tshem tawm cov primers, thiab synthesis ntawm DNA. Lub tshuab thermocycler yog siv los ua cov tshuaj tiv thaiv no kom nws tuaj yeem hloov kho qhov kub thiab txias sai thiab raug. Cov ntawv thov ntawm PCR yog kev tshawb nrhiav txhaum cai, DNA ntiv tes, tshawb pom cov kab mob, thiab kev tshuaj xyuas DNA ntawm tib neeg hom thaum ntxov.
Cov PCR yog dab tsi?
Muaj peb theem loj ntawm cov pa PCR, uas yog; DNA amplification theem, sib cais ntawm PCR, thiab nrhiav kom tau cov khoom. Kev sib cais ntawm DNA ntu feem ntau yog ua los ntawm agarose gel electrophoresis. Tom qab ntawd cov khoom yog stained nrog etheiduim bromide. Thaum kawg, kev kuaj pom tau tiav los ntawm kev pom ntawm cov hlua mus rau gels hauv qab UV teeb. Yog li ntawd, qhov kawg tshwm sim ntawm cov pa PCR tsis tau qhia raws li tus lej. Feem ntau cov pa PCR tsuas tuaj yeem ntes tau ib qho kev ntsuas xwb.
PCR tiag tiag yog dab tsi?
Lub sijhawm tiag tiag PCR tuaj yeem kuaj xyuas qhov kev nthuav dav ntawm cov khoom lag luam, raws li cov khoom sib txuas. Nrog rau kev txhim kho thev naus laus zis, PCR tau dhau los ua cov txheej txheem nrov heev, tshwj xeeb tshaj yog rau kev tshawb nrhiav thiab txheeb xyuas cov kab mob hauv cov khoom noj. Lub sijhawm tiag tiag PCR siv lub florescent dye system thiab thermocycler nruab nrog fluorescent- kuaj pom muaj peev xwm.
Qhov txawv ntawm Pa PCR thiab Real-time PCR yog dab tsi?
• Pa PCR siv sijhawm ntau dua vim nws siv gel electrophoresis los tshuaj xyuas cov khoom lag luam PCR amplified. Hauv qhov sib piv, PCR lub sijhawm tiag tiag siv sijhawm tsawg dua vim tias nws tuaj yeem kuaj pom kev nthuav dav thaum lub sijhawm ntxov ntawm cov tshuaj tiv thaiv.
• PCR lub sijhawm tiag tiag sau cov ntaub ntawv ntawm theem kev loj hlob ntawm PCR thaum ib txwm PCR sau cov ntaub ntawv ntawm qhov kawg ntawm cov tshuaj tiv thaiv.
• Cov ntsiab lus kawg ntawm PCR cov qauv yuav tsis yog qhov tseeb, tab sis cov txiaj ntsig ntawm PCR lub sijhawm tiag tiag yog qhov tseeb heev.
• PCR lub sijhawm tiag tiag yog qhov nkag siab dua li PCR cov qauv.
• Pa PCR muaj kev daws teeb meem tsis zoo thaum lub sijhawm PCR tuaj yeem pom qhov hloov pauv me me vim qhov kev daws teeb meem siab.
• Xaus point nrhiav pom ntawm cov pa PCR muaj luv luv dynamic ntau thaum lub sij hawm real-time PCR nrhiav tau dav dynamic ntau.
• Tsis zoo li cov qauv PCR, cov txheej txheem kev kuaj pom muaj nyob hauv lub sijhawm PCR.
• Pa PCR yog qhov ntse heev thiab siv zog ntau dua li PCR lub sijhawm.
• Tsis zoo li PCR lub sijhawm tiag tiag, PCR cov qauv tsis tuaj yeem cais tawm ntawm cov kab mob tuag thiab muaj sia nyob.
• PCR Real-time siv cov fluorescent dye system los xyuas cov khoom thaum cov khoom siv PCR siv ethidium bromide thiab UV teeb kom pom cov bands hauv agarose gel nruab nrab.
