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I-Foreasy Taq DNA Polymerase

I-Foreasy Taq DNA Polymerase Isithombe Esifakiwe
  • I-Foreasy Taq DNA Polymerase

Incazelo yekhithi:

◮Ukucaciswa okuphezulu: I-enzyme inomsebenzi othile wokuqalisa ukushisa.

◮Ukukhulisa Ngokushesha: 10 sec/kb.

◮Isifanekiso esizivumelanisa nezimo kakhulu: singasetshenziswa ukuze kukhuliswe kahle inani eliphakeme le-GC, izifanekiso ezihlukahlukene okunzima ukuzikhulisa ze-DNA.

◮Ukuthembeka okuqinile: I-Taq Enzyme evamile izikhathi ezi-6.

◮Ukuzinza okuqinile kokushisa: Ingabekwa ku-37 °C isonto lonke futhi igcine umsebenzi ongaphezu kuka-90%.

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Imininingwane Yomkhiqizo

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FAQ

Incazelo

I-Foreasy Taq DNA Polymerase iyi-enzyme entsha ye-Taq evezwa kubhaktheriya yobunjiniyela be-Escherichia coli ngobuchwepheshe bokuhlanganiswa kabusha kwezakhi zofuzo.I-enzyme ngokwayo inomsebenzi othile wokuqalisa ukushisa futhi ingasetshenziselwa i-PCR ne-qPCR evamile;inomsebenzi we-DNA polymerase ongu-5'→3' kanye nomsebenzi we-exonuclease ongu-5'→3', kodwa akukho 3'→5' umsebenzi we-exonuclease.

Izingxenye zekhithi

Isakhi

 I-IM-01011 I-IM-01012 I-IM-01013
I-Foreasy Taq DNA Polymerase(5 U/μL)  5000 U (1 mL)  50 KU (10 mL)  500 KU (100 mL)
2× I-Taq Reaction Buffer  25 mL × 5  250 mL × 5  500 ml × 25

Izici&izinzuzo

- Ukucaciswa okuphezulu: I-enzyme inomsebenzi othile wokuqalisa ukushisa.

- Ukukhulisa Ngokushesha: 10 sec/kb.

- Isifanekiso esizivumelanisa nezimo kakhulu: singasetshenziswa ukuze kukhuliswe kahle inani eliphakeme le-GC, izifanekiso ezihlukahlukene ze-DNA okunzima ukuzikhulisa.

- Ukuthembeka okuqinile: I-Taq Enzyme ejwayelekile izikhathi ezi-6.

- Ukuqina okushisayo okuqinile: Ingabekwa ku-37 ° C isonto lonke futhi igcine umsebenzi ongaphezu kuka-90%

Isicelo sekhithi

Izinhlelo ezihlukahlukene ze-PCR/qPCR kanye nezinhlelo ze-PCR eziqondile

I-PCR yokukhulisa izingcezu ze-DNA

Ukulebula kwe-DNA

Ukulandelana kwe-DNA

I-PCR A-tailed

U Incazelo

I-1U: Inani le-enzyme edingekayo ukuze kufakwe i-10 nmol ye-deoxynucleotides entweni engancibiliki i-asidi kusetshenziswa i-DNA yesidoda se-salmon ecushiwe njengesifanekiso/i-primer imizuzu engu-30 ku-74°C.

Isimo Sokusabela

Izinga lokushisa Isikhathi Umjikelezo
37°C 5 imiz 1
94°C 5 imiz 1
94°C 10 amasekhondi  

35
60°C 10 amasekhondi
72°C 20 isekhondi/kb
72°C 2 imiz 1

Isitoreji

-20 ± 5 °C iminyaka emi-2 noma ku -80 °C ukuze igcinwe isikhathi eside.

  • Okwedlule:
  • Olandelayo:

    • Awekho amasiginali wokukhulisa umsindo

    1.I-Taq DNA Polymerase kukhithi ilahlekelwa umsebenzi wayo ngenxa yokugcinwa okungalungile noma ukuphelelwa yisikhathi kwekhithi.
    Okutuswayo: Qinisekisa izimo zokugcinwa kwekhithi;wengeze kabusha inani elifanele le-Taq DNA Polymerase ohlelweni lwe-PCR noma uthenge Ikhithi ye-PCR Yesikhathi Sangempela entsha ngokuhlolwa okuhlobene.

    2.Kunenqwaba yama-inhibitors e-Taq DNA Polymerase kusifanekiso se-DNA.
    Isiphakamiso: Hlanza kabusha isifanekiso noma wehlise inani lesifanekiso esisetshenzisiwe.

    3.I-Mg2+ concentration ayifanele.
    Isincomo: Ukugxiliswa kwe-Mg2+ kwe-2× Real PCR Mix esikunikezayo ngu-3.5mM.Kodwa-ke, kwezinye iziqalo ezikhethekile nezifanekiso, ukugxila kwe-Mg2+ kungase kube phezulu.Ngakho-ke, ungakwazi ukwengeza i-MgCl2 ngokuqondile ukuze uthuthukise ukugxila kwe-Mg2+.Kunconywa ukuthi ukhuphule i-Mg2+ 0.5mM isikhathi ngasinye ukuze kuthuthukiswe.

    I-4.Izimo zokukhulisa i-PCR azifaneleki, futhi ukulandelana kwe-primer noma ukugxila akufanelekile.
    Isiphakamiso: qinisekisa ukunemba kokulandelana kwe-primer futhi i-primer ayizange yehliswe;uma isignali yokukhulisa i-amplification ingalungile, zama ukwehlisa izinga lokushisa le-anneal futhi ulungise ukugxila kwe-primer ngendlela efanele.

    5.Inani lesifanekiso lincane kakhulu noma liningi kakhulu.
    Okunconyiwe: Yenza isifanekiso sokuhlanjululwa kwe-gradient yomugqa, bese ukhetha ukugxiliswa kwesifanekiso esinomphumela we-PCR ongcono kakhulu wokuhlolwa kwe-PCR ye-Real Time.

    I-NTC inevelu ye-fluorescence ephezulu kakhulu

    Ukungcola kwe-1.Reagent okubangelwa ngesikhathi sokusebenza.
    Okunconyiwe: Faka esikhundleni ngama-reagents amasha okuhlolwa kwe-PCR ye-Real Time.

    2.Ukungcola kwenzeke ngesikhathi sokulungiswa kohlelo lokusabela kwe-PCR.
    Okunconyiwe: Thatha izinyathelo zokuzivikela ezidingekayo ngesikhathi sokusebenza, njengokuthi: ukugqoka amagilavu ​​e-latex, ukusebenzisa ithiphu ye-pipette ngesihlungi, njll.

    3.Iziqalo zehlisiwe, futhi ukucekelwa phansi kweziqalo kuzodala ukukhuliswa okungaqondile.
    Isiphakamiso: Sebenzisa i-SDS-PAGE electrophoresis ukuze uthole ukuthi iziqalo zonakalisiwe, futhi esikhundleni sazo ufake iziqalisi ezintsha zokuhlolwa kwe-PCR ye-Real Time.

    I-Primer dimer noma ukukhulisa okungaqondile

    1.I-Mg2+ concentration ayifanele.
    Okutuswayo: Ukugxiliswa kwe-Mg2+ ye-2× Real PCR EasyTM Mix esiyinikezayo ngu-3.5 mM.Kodwa-ke, kwezinye iziqalo ezikhethekile nezifanekiso, ukugxila kwe-Mg2+ kungase kube phezulu.Ngakho-ke, ungakwazi ukwengeza i-MgCl2 ngokuqondile ukuze uthuthukise ukugxila kwe-Mg2+.Kunconywa ukuthi ukhuphule i-Mg2+ 0.5mM isikhathi ngasinye ukuze kuthuthukiswe.

    2.Izinga lokushisa le-PCR annealing liphansi kakhulu.
    Isiphakamiso: Nyusa izinga lokushisa le-PCR le-anneal ngo-1℃ noma ngo-2℃ isikhathi ngasinye.

    3.Umkhiqizo we-PCR mude kakhulu.
    Okunconyiwe: Ubude bomkhiqizo we-Real Time PCR kufanele bube phakathi kuka-100-150bp, bungabi ngaphezu kuka-500bp.

    I-4.Ama-primers awonakala, futhi ukuchithwa kwama-primers kuzoholela ekubukeni kokukhulisa okuqondile.
    Isiphakamiso: Sebenzisa i-SDS-PAGE electrophoresis ukuze uthole ukuthi iziqalo zonakalisiwe, futhi esikhundleni sazo ufake iziqalisi ezintsha zokuhlolwa kwe-PCR ye-Real Time.

    5.Isistimu ye-PCR ayilungile, noma isistimu incane kakhulu.
    Isiphakamiso: Isistimu yokusabela ye-PCR incane kakhulu izobangela ukunemba kokutholwa kwehle.Kungcono kakhulu ukusebenzisa isistimu yokusabela enconywe ithuluzi le-PCR lobuningi ukuze uqalise kabusha ukuhlolwa kwe-PCR ye-Real Time.

    Ukuphindaphinda okubuthakathaka kwamanani omthamo

    1.Ithuluzi alisebenzi kahle.
    Isiphakamiso: Kungase kube namaphutha phakathi kwembobo ngayinye ye-PCR yethuluzi, okuholela ekukhiqizeni kabusha okubi ngesikhathi sokuphathwa kwezinga lokushisa noma ukutholwa.Sicela uhlole ngokulandela imiyalelo yethuluzi elihambisanayo.

    2.Ukuhlanzeka kwesampula akukuhle.
    Okunconyiwe: Amasampuli angcolile azoholela ekukhiqizeni kabusha okubi kokuhlolwa, okufaka ukuhlanzeka kwesifanekiso nama-primers.Kungcono ukuhlanza kabusha isifanekiso, futhi ama-primer ahlanzwa kangcono yi-SDS-PAGE.

    3.Ukulungiswa kohlelo lwe-PCR nesikhathi sokugcina side kakhulu.
    Isiphakamiso: Sebenzisa isistimu ye-PCR ye-Real Time yesilingo se-PCR ngokushesha ngemva kokulungiselela, futhi ungayishiyi eceleni isikhathi eside kakhulu.

    I-4.Izimo zokukhulisa i-PCR azifaneleki, futhi ukulandelana kwe-primer noma ukugxila akufanelekile.
    Isiphakamiso: qinisekisa ukunemba kokulandelana kwe-primer futhi i-primer ayizange yehliswe;uma isignali yokukhulisa i-amplification ingalungile, zama ukwehlisa izinga lokushisa le-anneal futhi ulungise ukugxila kwe-primer ngendlela efanele.

    5.Isistimu ye-PCR ayilungile, noma isistimu incane kakhulu.
    Isiphakamiso: Isistimu yokusabela ye-PCR incane kakhulu izobangela ukunemba kokutholwa kwehle.Kungcono kakhulu ukusebenzisa isistimu yokusabela enconywe ithuluzi le-PCR lobuningi ukuze uqalise kabusha ukuhlolwa kwe-PCR ye-Real Time.

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