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Isikhathi Sangempela PCR Easyᵀᴹ-Taqman

Isikhathi Sangempela PCR Easyᵀᴹ-Taqman Isithombe Esifakiwe
  • Isikhathi Sangempela PCR Easyᵀᴹ-Taqman
  • Isikhathi Sangempela PCR Easyᵀᴹ-Taqman

Incazelo yekhithi:

◮I-Simple—2× PCR Mix ukuze kwehliswe iphutha lokuhlola nesikhathi sokusebenza

◮I-Specific-optimized buffer kanye ne-hot-start Taq enzyme ingavimbela ukukhulisa okungaqondile kanye nokwakheka kwe-primer dimer

◮Ukuzwela okuphezulu-kungathola amakhophi aphansi esifanekiso

◮Ukuguquguquka okuhle—kuhambisana nezinsimbi eziningi zesikhathi sangempela ze-PCR

amandla angaphambili


Imininingwane Yomkhiqizo

Omaka bomkhiqizo

FAQ

Izincazelo zekhithi

I-2X Real PCR EasyTMI-Mix-Taqman inikezwe i-Real Time PCR EasyTM-Ikhithi ye-Taqman iwuhlelo olusha lwe-premix olusebenzisa ama-fluorescent probe athile okusabela kokukhulisa i-Real Time PCR, okungathuthukisa kakhulu ukucaciswa komkhiqizo nokuzwela kokusabela.I-ROX inikezwa njengodayi wokulawula wangaphakathi.

2X Real PCR EasyTMI-Mix-Taqman iqukethe i-Taq DNA Polymerase eyingqayizivele ye-Foregene.Uma iqhathaniswa nama-enzyme e-Taq ajwayelekile, inezinzuzo zokusebenza kahle kokukhulisa i-amplification, ikhono elithile eliqinile lokukhulisa kanye nezinga lokungafani eliphansi.Inganciphisa ukukhulisa okungaqondile futhi ithuthukise ukunemba kwe-PCR.

Imininingwane

Isikhathi Sangempela PCR EasyTM-Taqaman

Ukwakhiwa kwekhithi (20μl system)

QP-01021

QP-01022

QP-01023

QP-01024

200T

500T

1000T

2000T

2×I-PCR yangempelaKululaTMHlanganisa-Taqaman

1 ml ×2

1.7 ml ×3

1.7 ml ×6

1.7 ml ×12

20×I-ROX Reference Dye

200 μl

0.5ml

1 ml

1 ml × 2

I-DNase-Free ddH2O

1.7 ml

1.7 ml ×2

10 ml

20 ml

Iumyalo

1

1

1

1

Izici&izinzuzo

■ I-Simple—2X PCR Mix ukuze kwehliswe iphutha lokuhlola nesikhathi sokusebenza

■ Ibhafa ecacisiwe—elungiselelwe kahle kanye ne-enzyme ye-Taq eqala ukushisa ingavimbela ukukhulisa okungaqondile kanye nokwakheka kwe-primer dimer.

■ Ukuzwela okuphezulu—kukwazi ukubona amakhophi aphansi esifanekiso

■ Ukuguquguquka okuhle—kuhambisana nezinsimbi eziningi zesikhathi sangempela ze-PCR

Isicelo sekhithi

Ukuhlaziywa kwe-qPCR

Ukugeleza komsebenzi

I-RT PCR-Taqman
Umfanekiso we-RT PCR-Taqman

Umfanekiso

Isitoreji nempilo yeshelufu

Le kit kufanele igcinwe kude nokukhanya futhi kufanele igcinwe ku -20 ℃.Uma isetshenziswa njalo, ingabuye igcinwe ku-4℃ isikhathi esifushane (izinsuku eziyi-10).

  • 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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