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I-Foreasy HS Taq DNA Polymerase

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

Incazelo yekhithi:

◮Ukucaciswa okuphezulu: I-enzyme enomsebenzi ophezulu wokuqalisa ukushisa.

◮Ukukhulisa Ngokushesha: 10 sec/kb.

◮Ukuvumelana nezimo kwesifanekiso esiphezulu : kungasetshenziswa ukukhulisa kahle OkuphezuluGCinanifuthiizifanekiso ezihlukahlukene ze-DNA okunzima ukuzikhulisa.

◮Ukwethembeka okuqinile: Ukwethembeka izikhathi ezingu-6of i-Taq enzyme ejwayelekile.

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Imininingwane Yomkhiqizo

Omaka bomkhiqizo

FAQ

Incazelo

I-Foreasy HS Taq DNA Polymerase iyi-enzyme entsha ye-Taq evezwa kubhaktheriya yobunjiniyela be-Escherichia coli ngobuchwepheshe bokuhlanganiswa kabusha kwezakhi zofuzo.Ngemuva kokuthi i-enzyme iphathwe ngenqubo ekhethekile, yona's umsebenzi uyavinjwa ngaphambi kokwenza kusebenze okushisayo, ngaleyo ndlela kuvimbela ukukhulisa okungaqondile okubangelwa ukuhunyushwa okungaqondile kwama-primer noma ama-primer dimers ngaphansi kwezimo eziphansi lokushisa.Lo mkhiqizo ulungele i-PCR React ethileion, M ultiple x PCR , okuqukethwe okuphezulu kwe-GC ( > 60%) ,ngeisakhiwo sesibilinoma okunyei-genom yangemuva eqinileicsi-amplification kanye ne-genom enkuluicsukutholwa kwe-amplification.I-enzyme inomsebenzi ongu-5' → 3' we-DNA polymerase kanye nomsebenzi ongu-5' → 3' we-exonuclease, kodwa akukho 3' → 5' umsebenzi we-exonuclease.

Izingxenye zekhithi

Isakhi I-IM-01021 I-IM-01022 I-IM-01023
I-Foreasy HS Taq DNA Polymerase (5 U/μL)  5000 U (1 mL)  50 KU (10 mL)  500 KU (100 mL)
2× I-Taq Reaction Buffer  25mL × 5  250 mL × 5  500 mL × 25

Izici&izinzuzo

- Ukucaciswa okuphezulu: I-enzyme enomsebenzi ophezulu wokuqalisa ukushisa.

- Ukukhulisa Ngokushesha: 10 sec/kb.

- Ukuguquguquka kwesifanekiso esiphezulu : kungasetshenziswa ukukhulisa kahle OkuphezuluGCinanifuthiizifanekiso ezihlukahlukene ze-DNA okunzima ukuzikhulisa.

- Ukuthembeka okuqinile: Ukwethembeka izikhathi eziyisi-6of i-Taq enzyme ejwayelekile.

Isicelo sekhithi

- Uhlelo oluhlukahlukene lwe-PCR/qPCR kanye nohlelo lwe-PCR oluqondile

- I-PCR Amplified DNA Fragment

- I-DNA mark

- Ukulandelana kwe-DNA

- PCR plus A umsila

Incazelo Yomsebenzi

I-1U : Inani le-enzyme edingekayo ukuze kufakwe i-10 nmol yeI-DNAodabeni olungancibiliki nge-asidi kusetshenziswa i-DNA yesidoda se-salmon esenziwe yasebenza njengesifanekiso / isiqalisi, 74 °C, imizuzu engama-30.

Isimo Sokusabela

Izinga lokushisa Isikhathi sokuphendula Isikhathi somjikelezo
37°C 5 imiz 1
94°C 5 imiz 1
94°C 10 amasekhondi  40
60°C 10 amasekhondi

Qaphela:Kumasistimu angu-10 µL no-20 µL, engeza ivolumu elinganayo yamafutha amaminerali uma umjikelezo oshisayo ungenaso isivalo sokushisa .

Izimo zokusabela kwe-PCR ziyahlukahluka kuye ngezimo zesakhiwo sezifanekiso, iziqalo, nokunye okunjalo.Emsebenzini othize, kuyadingeka ukuklama izimo zokusabela ezifanele, okuhlanganisa izinga lokushisa le-annealing, isikhathi sokunwetshwa, njll., ngokuya ngezimo ezithile ezifana nohlobo lwesifanekiso, usayizi wocezu oluqondiwe, ukulandelana kwesisekelo kwesiqephu esikhulisiwe, nokuqukethwe kwe-GC nobude be-primer.

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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