Qhov sib txawv tseem ceeb ntawm cov xov tooj cua thiab cov xov tooj cua tsis yog cov xov tooj cua yog cov xov tooj cua yog ib leeg-kab DNA lossis RNA ib ntus uas tau sau nrog cov isotopes radioactive thaum cov kev sojntsuam uas tsis yog-radioactive yog ib leeg-stranded DNA lossis RNA sequences uas tau sau nrog cov tshuaj lossis fluorescent tag.
Nucleic acid hybridization yog cov txheej txheem tseem ceeb hauv molecular biology, tshwj xeeb tshaj yog hauv kev kuaj mob microbial. Nws pab txheeb xyuas lossis txheeb xyuas cov nucleic acid ib ntus. Nyob rau hauv cov txheej txheem no, nucleic acids yog tsau rau ib tug khoom nto thiab hybridized nrog ib tug sojntsuam. Ib qho kev sojntsuam yog ib feem ntawm DNA lossis RNA uas yog ntxiv rau qhov kev txaus siab. Yog hais tias lub hom phiaj ua ntu zus nyob rau hauv cov qauv, qhov kev sojntsuam yuav hybridize nrog nws thiab ua kom nws ntes tau. Muaj ob hom kev sojntsuam xws li xov tooj cua thiab tsis muaj xov tooj cua. Yog li ntawd, peb tuaj yeem sau cov kev sojntsuam nrog cov xov tooj cua lossis cov fluorescent tag.
Dab tsi yog Radioactive Probes?
Xov tooj cua sojntsuam yog ib leeg-stranded DNA lossis RNA fragments nrog cov xov tooj cua. Radioisotopes yog siv los npaj cov tshuaj tiv thaiv kab mob. Radioisotopes 32P, 33P thiab 35S yog feem ntau siv rau hauv daim ntawv lo ntawm kev sojntsuam. Ntxiv mus, radioisotopes 3 H thiab 125 1 kuj tau siv rau qhov tsawg dua hauv kev sau npe ntawm kev sojntsuam. Tab sis lawv tau siv rau cov ntawv thov tshwj xeeb. Ntawm cov xov tooj cua sib txawv, 32 P yog cov isotope feem ntau siv hauv kev sau npe xov tooj cua.
Radioactive probes muab qib siab dua ntawm kev ntseeg siab thiab tshwj xeeb. Yog li ntawd, lawv muab siab tshaj plaws rhiab heev thiab tso cai kom muaj nuj nqis ntawm cov hom phiaj sib lawv liag. Txawm li cas los xij, muaj ntau qhov tsis zoo cuam tshuam nrog kev sojntsuam xov tooj cua. Lawv muaj luv ib nrab neej. Ntxiv mus, lawv muaj kev phom sij thiab ntau lawm, siv thiab pov tseg yog teeb meem thaum tuav. Tsis tas li ntawd, kev npaj tshuaj tua kab mob radioactive yog ib txheej txheem kim heev. Yog li ntawd, vim muaj teeb meem kev nyab xeeb thiab tus nqi, cov xov tooj cua xov tooj cua tsis tau siv los ua cov kev sojntsuam tsis muaj xov tooj cua niaj hnub no.
Nonradioactive Probes yog dab tsi?
Nonradioactive probes yog hom thib ob ntawm kev sojntsuam uas tau sau tshuaj. Digoxigenin yog ib qho kev sojntsuam uas tsis yog-radioactive, uas yog ib qho tshuaj tiv thaiv kab mob. Digoxigenin probes yog qhov tshwj xeeb thiab rhiab heev. Biotin yog lwm daim ntawv lo siv hauv kev tshuaj ntsuam xyuas uas tsis yog-radioactive. Biotin / Streptavidin thiab Digoxigenin / Antibody-detection systems yog cov feem ntau siv cov tshuaj tiv thaiv tsis muaj zog hauv hybridization. Tsis tas li ntawd, horseradish peroxidase system yog lwm qhov kev sojntsuam uas tsis yog-radioactive. Thaum cov kev sojntsuam uas tsis yog-radioactive no yog hybridized nrog lub hom phiaj sib lawv liag, lawv tuaj yeem kuaj pom ntawm autoradiography lossis lwm yam kev siv duab.
Daim duab 01: Kev sib xyaw nrog Nonradioactive Probes
Nonradioactive probes tau siv ntau zaus hauv nucleic acid hybridization dua li xov tooj cua. Qhov no yog vim hais tias cov kev sojntsuam nonradioactive tsis cuam tshuam nrog cov khoom siv phom sij. Tsis tas li ntawd, txoj kev tshawb nrhiav tsis muaj hluav taws xob xav tau lub sij hawm raug luv dua kom pom cov teeb liab hybridization. Txawm li cas los xij, cov kauj ruam koom nrog hauv DNA hybridization nrog cov kev sojntsuam uas tsis yog radioactive feem ntau yog qhov nyuaj thiab siv sijhawm. Tsis tas li ntawd, cov kev daws teeb meem uas muag muag yog kim.
Dab tsi zoo sib xws ntawm Radioactive thiab Nonradioactive Probes?
- Radioactive thiab nonradioactive probes yog ob hom kev sojntsuam siv hauv nucleic acid hybridization.
- Lawv pab txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau ua tus qauv.
- Ob hom kev sojntsuam yog qhov sib luag thiab tshwj xeeb.
Qhov txawv ntawm Radioactive thiab Nonradioactive Probes yog dab tsi?
Radioactive probes yog ib leeg-stranded DNA lossis RNA sequences sau nrog radioactive isotopes, thaum nonradioactive probes yog ib leeg-stranded DNA los yog RNA sequences sau nrog ib tug tshuaj tag. Yog li, qhov no yog qhov sib txawv tseem ceeb ntawm cov xov tooj cua thiab tsis muaj xov tooj cua. Tsis tas li ntawd, radioactive isotopes yog qhov txaus ntshai. Yog li ntawd, cov kev sojntsuam xov tooj cua yog qhov txaus ntshai heev, thaum cov kev sojntsuam uas tsis yog-radioactive tsis muaj kev phom sij.
Ntxiv mus, lwm qhov sib txawv ntawm cov xov tooj cua thiab cov xov tooj cua tsis zoo yog lawv qhov tsis zoo. Ib nrab-lub neej luv luv thiab cov teeb meem cuam tshuam nrog lawv cov khoom tsim, siv thiab pov tseg yog qhov tsis zoo ntawm kev siv xov tooj cua sojntsuam. Ntawm qhov tod tes, cov kauj ruam koom nrog hauv DNA hybridization nrog kev sojntsuam uas tsis yog-radioactive feem ntau yog tedious thiab siv sijhawm.
Hauv qab infographic qhia ntau qhov kev sib piv ntsig txog qhov sib txawv ntawm cov xov tooj cua thiab tsis muaj xov tooj cua.
Summary – Radioactive vs Nonradioactive Probes
Ib qho kev sojntsuam yog ib feem ntawm DNA lossis RNA uas muaj cov nucleotide ib ntus uas yog ntxiv rau qhov kev txaus siab. Txhawm rau txhawm rau txhawm rau ua ntu zus, kev soj ntsuam tuaj yeem sau npe tuaj yeem sau npe tuaj yeem sau npe, fluorescently lossis tshuaj. Kev sojntsuam khi nrog cov kab ke sib luag hauv cov qauv. Radioactive probes yog sau nrog cov isotopes radioactive thaum uas tsis yog-radioactive probes yog sau nrog biotin, digoxigenin los yog horseradish peroxidase. Yog li, qhov no yog qhov sib txawv tseem ceeb ntawm cov xov tooj cua thiab cov xov tooj cua tsis zoo.
