Qhov Tseem Ceeb Qhov txawv - RAPD vs RFLP
Cov cim caj ces yog siv hauv Molecular Biology txhawm rau txheeb xyuas cov caj ces txawv ntawm tib neeg thiab hom. Random Amplified Polymorphic DNA (RAPD) thiab Restriction Fragment Length Polymorphism (RFLP) yog ob lub cim tseem ceeb molecular uas niaj hnub siv hauv chav kuaj. RAPD yog ua nrog luv luv thiab arbitrary oligonucleotide primers, thiab nws yog raws li nyob rau hauv random amplification ntawm ntau qhov chaw thoob plaws hauv lub template DNA ntawm lub cev. RFLP yog ua nrog ib qho kev txwv tshwj xeeb endonuclease, thiab nws yog raws li polymorphism ntawm qhov tshwm sim txwv tsis pub tawg thiab hybridization. Qhov sib txawv tseem ceeb ntawm RAPL thiab RFLP yog tias RAPD yog ib hom PCR txheej txheem ua yam tsis muaj kev paub ua ntej thaum RFLP tsis koom nrog PCR thiab yuav tsum muaj kev paub ua ntej los ua cov txheej txheem.
RAPD yog dab tsi?
RAPD yog ib qho txiaj ntsig molecular cim hauv molecular biology. Nws yog cov txheej txheem ceev thiab yooj yim. RAPD tuaj yeem txhais tau tias yog ib txoj hauv kev uas ua rau polymorphic DNA sequences raws li qhov tshwm sim ntawm kev nthuav dav ntawm ntau qhov chaw ntawm lub hom phiaj DNA qauv. RAPD siv luv oligonucleotide primers nrog arbitrary sequences rau PCR amplification. Primers yog cov khoom tsim tawm yam tsis muaj kev paub ua ntej. Li no, nws suav hais tias yog ib qho yooj yim thiab siv tau.
Cov kauj ruam tseem ceeb hauv qab no yog koom nrog RAPD.
- Kev rho tawm ntawm lub hom phiaj DNA
- Kev nthuav dav ntawm ntau qhov chaw ntawm lub hom phiaj DNA siv cov txheej txheem xaiv xaiv
- Gel electrophoresis ntawm amplified PCR khoom
- Staining nrog ethidium bromide thiab txheeb xyuas cov polymorphism
Raws li qhov sib txawv ntawm qhov sib txawv ntawm cov primer annealing, cov khoom sib txawv nrog qhov ntev sib txawv yog tsim thaum lub sijhawm nthuav dav. Li no, banding qauv ntawm cov gels txawv ntawm cov tib neeg thiab hom. Yog li, RAPD ua rau kev tshawb pom ntawm kev hloov pauv caj ces ntawm cov kab mob hauv kev txheeb xyuas thiab sib txawv.
RAPD yog siv rau hauv ntau yam kev tshawb fawb ntawm molecular biology xws li kev txheeb xyuas qhov sib txawv ntawm cov noob caj noob ces ntawm cov tsiaj uas muaj feem xyuam nrog, gene mapping, DNA fingerprinting, txheeb xyuas cov kab mob qub, thiab lwm yam.
Daim duab 01: RAPD
RFLP yog dab tsi?
Txoj Kev Txwv Fragment Length Polymorphisms (RFLPs) yog cov cim molecular siv hauv molecular biology rau kev txheeb xyuas cov caj ces txawv hauv homologous DNA sequences. Nws yog thawj tus cim caj ces tsim rau DNA ntiv tes. Txhua yam kab mob tsim cov DNA tshwj xeeb thaum muaj kev txwv tshwj xeeb txwv enzymes. RFLP yog ib qho cuab yeej tseem ceeb hauv kev tsim cov DNA tshwj xeeb ntawm cov tib neeg thiab tshawb nrhiav cov kev hloov pauv caj ces ntawm lawv. Thaum cov qauv DNA tau zom nrog cov kev txwv tshwj xeeb endonucleases, nws muab cov DNA sib txawv uas yog qhov tshwj xeeb rau txhua tus neeg. Yog li ntawd, lub hauv paus tseem ceeb ntawm txoj kev no yog kev tshawb nrhiav kev hloov pauv caj ces ntawm cov kab mob los ntawm kev txwv homologous DNA nrog cov kev txwv tshwj xeeb enzymes thiab kev tsom xam ntawm qhov ntev ntawm polymorphism ntawm gel electrophoresis thiab blotting. Blotting qauv yog tshwj xeeb rau txhua yam kab mob thiab ua tus yam ntxwv ntawm genotypes.
Cov kauj ruam nram qab no koom nrog RFLP.
- Kev cais cov DNA txaus los ntawm cov qauv
- Kev tawg ntawm DNA cov qauv nrog kev txwv tshwj xeeb endonucleases rau hauv ntu luv
- Kev sib cais ntawm qhov tshwm sim ntawm qhov sib txawv ntawm qhov ntev los ntawm agarose gel electrophoresis.
- Hloov cov gel profile rau hauv daim nyias nyias los ntawm Southern blotting
- Hybridization ntawm daim nyias nyias nrog cov ntawv lo sojntsuam thiab kev tshuaj xyuas ntawm cov ntu ntev polymorphism hauv txhua qhov profile
RFLP muaj ntau yam kev siv xws li kev kuaj mob ntawm cov kab mob qub txeeg qub teg, genome mapping, kev txheeb xyuas kev ua txhaum cai hauv kev tshawb fawb forensic, kuaj txiv neej, thiab lwm yam.
Daim duab 02: RFLP genotyping
Qhov txawv ntawm RAPD thiab RFLP yog dab tsi?
RAPD vs RFLP |
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| RAPD yog tus cim molecular raws li cov primers random thiab PCR. | RFLP yog tus cim molecular raws li kev tsim tawm ntawm qhov sib txawv ntawm qhov txwv qhov ntev. |
| Yuav tsum tau ua qauv | |
| Cov qauv DNA me me txaus rau kev tshuaj xyuas RAPD. | Ntau cov qauv DNA rho tawm yog xav tau rau kev tshuaj xyuas RFLP. |
| RAPD yog txheej txheem ceev. | RFLP yog cov txheej txheem siv sijhawm. |
| Primer Use | |
| Random primers yog siv thiab tib primers siv tau rau ntau hom. | Cov tshuaj tshwj xeeb tshwj xeeb yog siv hauv RFLP rau kev sib xyaw. |
| Kev ntseeg ntawm cov txheej txheem tsawg dua piv rau RFLP. | RFLP yog cov txheej txheem txhim khu kev qha. |
| RAPD cuam tshuam rau yav qab teb blotting. | Southern blotting yog ib kauj ruam ntawm RFLP. |
| Kev Tshawb Fawb Txog Kev Sib Tw Allelic | |
| Allelic variations tsis tuaj yeem kuaj pom los ntawm RAPD. | Alllelic variations tuaj yeem kuaj pom los ntawm RFLP. |
| Nyob rau hauv Kev Paub | |
| RAPD tsis xav tau kev paub ua ntej. | Kev paub ua ntej ua ntej yog xav tau rau kev sojntsuam tsim. |
| PCR | |
| PCR koom nrog RAPD | PCR tsis koom nrog RFLP. |
| Reproducibility | |
| RAPD muaj qhov tsim tawm qis dua | RFLP muaj qhov ua tau zoo dua qub piv rau RAPD. |
Summary – RAPD vs RFLP
RAPD thiab RFLP yog cov cim tseem ceeb siv hauv molecular biology. Ob txoj kev muaj peev xwm txheeb xyuas qhov kev hloov pauv caj ces ntawm cov kab mob. RAPD yog ua los ntawm random primers. RFLP yog ua los ntawm kev txwv cov enzymes tshwj xeeb. Ob txoj hauv kev tsim DNA profile tshwj xeeb rau tib neeg cov kab mob. RAPD tau koom nrog ob peb kauj ruam dua li RFLP. Tab sis nws tsim cov txiaj ntsig tsis tshua muaj kev ntseeg siab thiab rov tsim dua li RFLP. Qhov no yog qhov txawv ntawm RAPD thiab RFLP.
