Qhov sib txawv tseem ceeb ntawm kev sib txuas thiab adapter yog qhov txuas tsis muaj qhov sib txuas thaum lub adapter muaj qhov sib txuas kawg.
DNA ligation yog txheej txheem ntawm kev koom nrog ob DNA molecules ua ke, tsim phosphodiester bonds. Lub enzyme hu ua DNA ligase catalyzes cov tshuaj tiv thaiv no. Nws yog ib qho ntawm cov kauj ruam tseem ceeb hauv niaj hnub molecular biological teb xws li recombinant DNA technology thiab DNA cloning. Lub ligation efficiency nyob ntawm qhov kawg ntawm DNA molecules yuav ligated. Muaj ob hom DNA xaus li nplaum kawg thiab blunt kawg. Ligation efficiency yog siab nrog cov nplaum kawg dua li nrog blunt kawg. Yog hais tias lub hom phiaj DNA molecules muaj blunt kawg, molecules hu ua adapters los yog linkers yuav pab tau. Adapters thiab linkers yog tshuaj synthesized oligonucleotide molecules uas pab hauv DNA ligation. Lawv muaj qhov chaw txwv sab hauv thiab. Adapter muaj ib qhov nplaum kawg thiab ib qho kawg, thaum txuas txuas muaj ob qhov kawg.
Dab tsi yog Linker?
Linker yog cov tshuaj sib xyaw ua ke ntawm oligonucleotide uas yog ob txoj hlua khi. Linker muaj ob qhov kawg. Linker yog siv los ligate DNA molecules uas muaj blunt kawg rau vectors. Nws muaj ib lossis ntau qhov chaw txwv sab hauv. Cov chaw txwv no ua haujlwm raws li qhov chaw paub txog kev txwv enzymes.
Daim duab 01: Linker
Tom qab ligation, DNA raug txwv dua nrog kev txwv cov enzymes los tsim cov khoom sib txuas. EcoRI-linkers thiab sal-I linkers feem ntau siv txuas.
Dab tsi yog Adaptor?
Ib qho Adapter yog ob txoj hlua khi oligonucleotide siv los txuas ob DNA molecules ua ke. Nws yog ib ntus luv luv nrog ib qho kawg kawg thiab ib qho nplaum los yog cohesive kawg. Yog li ntawd, nws muaj ib leeg-stranded tus Tsov tus tw ntawm ib kawg, uas txhim khu kev ua tau zoo ntawm DNA ligation.
Daim duab 02: DNA Ligation los ntawm tus Adaptor
Ntxiv mus, lub adapter muaj qhov chaw txwv sab hauv. Yog li ntawd, tom qab ligation, DNA tuaj yeem raug txwv nrog kev txwv tsis pub tsim nyog enzymes thiaj li tsim tau ib qho tshiab protruding terminus. Ib qho tsis zoo ntawm adapters yog tias ob lub adapters tuaj yeem tsim dimmers los ntawm kev sib koom ua ke nrog lawv tus kheej. Qhov no tuaj yeem zam tau los ntawm kev kho lawv nrog cov enzyme hu ua alkaline phosphatase.
Dab tsi zoo sib xws ntawm Linker thiab Adaptor?
- Ob qhov txuas thiab adapter yog ob-stranded luv oligonucleotide sequences.
- Lawv nqa qhov chaw txwv sab hauv.
- Ntxiv mus, lawv yog cov tshuaj lom neeg DNA molecules thiab yog hluavtaws molecules.
- Lawv tuaj yeem txuas ob DNA molecules ua ke.
- Tom qab ligation ntawm linkers thiab adaptors, cov DNA tau raug txwv dua nrog kev txwv enzymes txhawm rau tsim cov nplaum kawg.
Qhov txawv ntawm Linker thiab Adaptor yog dab tsi?
A linker yog cov tshuaj sib xyaw ua ke luv luv oligonucleotide duplex nrog ob qhov kawg. Ib qho adapter yog cov tshuaj sib xyaw ua ke luv luv oligonucleotide duplex nrog ib qho nplaum kawg thiab ib qho kawg. Yog li, qhov no yog qhov sib txawv tseem ceeb ntawm kev sib txuas thiab adaptor. Ntxiv mus, adapters tuaj yeem tsim dimers, thaum cov txuas tsis tsim dimers. Yog li, qhov no yog qhov sib txawv tseem ceeb ntawm kev sib txuas thiab adaptor.
Hauv qab no yog cov ntsiab lus ntawm qhov sib txawv ntawm qhov txuas thiab adapter hauv daim ntawv tabular.
Summary – Linker vs Adaptor
Linker thiab adapter yog ob hom tshuaj sib xyaw ua ke oligonucleotides uas muaj txiaj ntsig zoo hauv ligating blunt-end DNA. Linker muaj ob qhov xaus, thaum lub adapter muaj ib qho kawg thiab ib qho kawg. Yog li, qhov no yog qhov sib txawv tseem ceeb ntawm kev sib txuas thiab adaptor. Lawv yog ob-stranded molecules uas muaj qhov chaw txwv sab hauv. Lawv tau dav siv nyob rau hauv recombinant DNA technology thiab DNA cloning.
