Cishe wonke amalungu avela kubasebenzi bethu abakhulu abahola kahle bazisa izidingo zamakhasimende kanye nokuxhumana kwebhizinisi kwe-OEM/ODMI-China Taq Plus DNA Polymerase, Uma unentshisekelo kunoma yikuphi okuthengiswayo kwethu noma ufisa ukukhuluma ngokuthenga okuklanyelwe, kufanele uzizwe ukhululekile ngokuphelele ukusithola.
Cishe wonke amalungu avela kubasebenzi bethu abakhulu abahola kahle bazisa izidingo zamakhasimende kanye nokuxhumana kwebhizinisi ngakhoI-China Taq Plus DNA Polymerase, Sungula ubudlelwano bebhizinisi besikhathi eside futhi uwine nawo wonke amakhasimende ethu, yabelana ngempumelelo futhi ujabulele injabulo yokusabalalisa imikhiqizo yethu emhlabeni wonke ndawonye.Sithembe uzozuza okuningi.Sicela uzizwe ukhululekile ukusithinta ukuze uthole ulwazi olwengeziwe, sikuqinisekisa ngokunaka kwethu okuhle ngaso sonke isikhathi.

Imininingwane

50×20μl rxns, 200×20μl rxns, 1000×20μl rxns, 2000×20μl rxns

I-2X Real PCR EasyTM Mix-Taqman ehlinzekwa yi-Real Time PCR EasyTM-Taqman kit iwuhlelo olusha lwe-premix olusebenzisa ama-fluorescent probes athile okusabela kokukhulisa i-Real Time PCR, okungathuthukisa kakhulu ukucaciswa komkhiqizo nokuzwela kokusabela.I-ROX inikezwa njengodayi wokulawula wangaphakathi.

I-2X Real PCR EasyTM 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.

Izingxenye zekhithi

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

Umfanekiso

Isitoreji kanye Nempilo yeshelufu

Le kit kufanele igcinwe kude nokukhanya futhi kufanele igcinwe ku -20 ℃.Uma isetshenziswa njalo, ingabuye igcinwe ku-4℃ isikhathi esifushane (izinsuku ezingu-10).Cishe wonke amalungu avela kubasebenzi bethu abahola kahle abahola kahle bazisa izidingo zamakhasimende kanye nokuxhumana kwebhizinisi kwe-OEM/ODM China Taq Plus DNA Polymerase, Uma unentshisekelo kunoma yikuphi okuthengiswayo kwethu noma ufisa ukukhuluma ngokuthenga okulungiselelwe, kufanele uzizwe ukhululekile ngempela ukusibamba.
I-OEM/ODM China China Taq Plus DNA Polymerase, Sungula ubudlelwano bebhizinisi besikhathi eside newinile nawo wonke amakhasimende ethu, yabelana ngempumelelo futhi ujabulele injabulo yokusabalalisa imikhiqizo yethu emhlabeni wonke ndawonye.Sithembe uzozuza okuningi.Sicela uzizwe ukhululekile ukusithinta ukuze uthole ulwazi olwengeziwe, sikuqinisekisa ngokunaka kwethu okuhle ngaso sonke isikhathi.

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.