Ngokuvamile sisebenza njengabasebenzi ababambekayo siqinisekisa ukuthi sizokunikeza okuzuzisa kakhulu okuncomekayo kanye nenani elihle kakhulu lokuthengisa le-4X DNA Rna One-Step Rt-Qpcr Multiplex Master Mix Kit, Sizoqhubeka silwela ukukhulisa inkampani yethu futhi sinikeze imikhiqizo esezingeni elifanele ngamanani wamanani anolaka.Noma yimuphi umbuzo noma ukuphawula kwaziswa kakhulu.Khumbula ukusibamba ngokukhululeka.
Ngokuvamile sisebenza njengabasebenzi ababambekayo siqinisekisa ukuthi sizokunikeza inzuzo enhle kakhulu kanye nenani elihle kakhulu lokuthengisaI-China Taq DNA Polymerase ne-Qpcr, Ngeshabhu ethuthukisiwe, ithimba labaklami abaqeqeshiwe kanye nesistimu yokulawula ikhwalithi eqinile, esekelwe ekupheleni okumakwe njengokumaketha kwethu, imikhiqizo yethu ithengiswa ngokushesha ezimakethe zaseYurophu nezaseMelika ngemikhiqizo yethu efana nengaphansi kweDeniya, Qingsiya kanye neYisilanya.

Izincazelo zekhithi

I-2X Real PCR Easy^TMI-Mix-Taqman inikezwe i-Real Time PCR Easy^TM-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 Easy^TMI-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

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 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).

Ngokuvamile sisebenza njengabasebenzi ababambekayo siqinisekisa ukuthi sizokunikeza okuhle kakhulu okuzuzisa kakhulu kanye nenani elingcono kakhulu lokuthengisa le-OEM/ODM Supplier High Sensitivity One-Step Rt-Qpcr Multiplex Master Mix Kit, Sizoqhubeka silwela ukukhulisa inkampani yethu futhi sinikeze imikhiqizo esezingeni eliphezulu enamazinga entengo anolaka.Noma yimuphi umbuzo noma ukuphawula kwaziswa kakhulu.Khumbula ukusibamba ngokukhululeka.
Umhlinzeki we-OEM/ODMI-China Taq DNA Polymerase ne-Qpcr, Ngeshabhu ethuthukisiwe, ithimba labaklami abaqeqeshiwe kanye nesistimu yokulawula ikhwalithi eqinile, esekelwe ekupheleni okumakwe njengokumaketha kwethu, imikhiqizo yethu ithengiswa ngokushesha ezimakethe zaseYurophu nezaseMelika ngemikhiqizo yethu efana nengaphansi kweDeniya, Qingsiya kanye neYisilanya.

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.