Umvelisi weSingle Strip iMag-Rod Comb Nucleic Acid Test Ukubonwa kwezinto eziLahlwayo
Sinikezela ngamandla amangalisayo kumgangatho ophakamileyo kunye nophuculo, ukuthengisa, ukuthengisa imveliso kunye nokuthengisa kunye nentengiso kunye nenkqubo yoMvelisi weSingle Strip Mag-Rod Comb Nucleic Acid Test Disposable Consumables, Ishishini lethu linyanzelisa ukuqamba izinto ezintsha ukukhuthaza uphuhliso oluzinzileyo lwenkampani, kwaye sisenze sibe ngababoneleli abakumgangatho ophezulu basekhaya.
Sinikezela ngamandla amangalisayo kumgangatho ophezulu kunye nophuculo, ukuthengisa, ukuthengisa imveliso kunye nokuthengisa kunye nentengiso kunye nenkquboI-China Nucleic Acid kunye ne-Medical Consumables, Sijongene nokhuphiswano oluqatha lwemarike yehlabathi, siye sasungula isicwangciso-qhinga sokwakha uphawu saza sahlaziya umoya “wenkonzo ejolise ebantwini nethembekileyo”, ngenjongo yokufumana ukuqondwa kwihlabathi kunye nophuhliso oluzinzileyo.
Iinkcazelo Kit
I-2X yokwenyani PCR EasyTMI-Mix-Taqman enikezelwa yi-Real Time PCR EasyTM-Ikhithi ye-Taqman yinkqubo entsha ye-premix esebenzisa i-fluorescent probes ye-Real Time PCR reactions amplification, enokuphucula kakhulu imveliso ethile kunye novakalelo lokuphendula.I-ROX inikezelwa njengedayi yolawulo lwangaphakathi.
2X Real PCR EasyTMI-Mix-Taqman iqulethe i-Foregene eshushu-isiqalo esisodwa se-Taq DNA Polymerase.Xa kuthelekiswa nee-enzymes ze-Taq eziqhelekileyo, ineengenelo zokusebenza kwe-amplification ephezulu, isakhono esithile esinamandla sokukhulisa kunye nesantya esisezantsi sokungahambelani.Inokunciphisa ukukhuliswa okungangqaliyo kunye nokuphucula ukuchaneka kwe-PCR.
Iinkcukacha
Ixesha lokwenyani PCR EasyTM-Taqman | ||||
Ukwakhiwa kwekhithi (20μl inkqubo) | QP-01021 | QP-01022 | QP-01023 | QP-01024 |
200T | 500T | 1000T | 2000T | |
I-2 × I-PCR yokwenene elulaTMUmxube-Taqman | 1 ml × 2 | 1.7 ml × 3 | 1.7 ml × 6 | 1.7 ml × 12 |
I-20 × i-ROX ye-Reference Dye | 200 μl | 0.5 ml | 1 ml | 1 ml × 2 |
DNase-Free ddH2O | 1.7 ml | 1.7 ml × 2 | 10 ml | 20 ml |
Umyalelo | 1 | 1 | 1 | 1 |
Iimpawu&izinto eziluncedo
■ Ilula—I-2X PCR Mix ukunciphisa impazamo yovavanyo kunye nexesha lokusebenza
■ I-Specific-optimized buffer kunye ne-hot-start Taq enzyme inokuthintela ukukhulisa okungangqalanga kunye nokwakheka kwe-primer dimer.
■ Uvakalelo oluphezulu-lukwazi ukubona iikopi ezisezantsi zetemplate
■ Ukuguquguquka okuhle—okuhambelana nezixhobo ezininzi zePCR zexesha lokwenyani
Isicelo seKit
Uhlalutyo lwe-qPCR
Ukuhamba komsebenzi
Umzobo
Ukugcinwa kunye nobomi beshelufu
Le khithi kufuneka igcinwe kude nokukhanya kwaye kufuneka igcinwe ku -20 ℃.Ukuba isetyenziswe rhoqo, inokugcinwa kwi-4 ℃ ixesha elifutshane (iintsuku ezili-10).
Sinikezela ngamandla amangalisayo kumgangatho ophakamileyo kunye nophuculo, ukuthengisa, ukuthengisa imveliso kunye nokuthengisa kunye nentengiso kunye nenkqubo yoMvelisi weSingle Strip Mag-Rod Comb Nucleic Acid Test Disposable Consumables, Ishishini lethu linyanzelisa ukuqamba izinto ezintsha ukukhuthaza uphuhliso oluzinzileyo lwenkampani, kwaye sisenze sibe ngababoneleli abakumgangatho ophezulu basekhaya.
Umenzi weI-China Nucleic Acid kunye ne-Medical Consumables, Sijongene nokhuphiswano oluqatha lwemarike yehlabathi, siye sasungula isicwangciso-qhinga sokwakha uphawu saza sahlaziya umoya “wenkonzo ejolise ebantwini nethembekileyo”, ngenjongo yokufumana ukuqondwa kwihlabathi kunye nophuhliso oluzinzileyo.
Akukho miqondiso yokukhulisa
I-1.I-Taq DNA Polymerase kwikiti ilahlekelwa ngumsebenzi wayo ngenxa yokugcinwa okungafanelekanga okanye ukuphelelwa yisikhathi kwekiti.
Ingcebiso: Qinisekisa imiqathango yokugcina ikhithi;yongeza kwakhona imali efanelekileyo ye-Taq DNA Polymerase kwinkqubo ye-PCR okanye uthenge i-Real Time PCR Kit kwimifuniselo ehambelanayo.
2.Zininzi ii-inhibitors ze-Taq DNA Polymerase kwi-template ye-DNA.
Ingcebiso: Hlaziya itemplate okanye unciphise inani letemplate esetyenzisiweyo.
3.I-concentration ye-Mg2 + ayifanelekanga.
Isincomo: I-Mg2 + yoxinaniso lwe-2× Real PCR Mix esibonelela ngayo yi-3.5mM.Nangona kunjalo, kwezinye iiprimers ezikhethekileyo kunye neetemplates, ugxininiso lweMg2 + lunokuba phezulu.Ke ngoko, unokongeza ngokuthe ngqo i-MgCl2 ukongeza i-Mg2+ yoxinaniso.Kuyacetyiswa ukuba kwandiswe i-Mg2+ 0.5mM ixesha ngalinye ukwenzela ukuba kuphuculwe.
4.Iimeko zokukhulisa i-PCR azifanelekanga, kwaye ukulandelelana kwe-primer okanye ukugxininiswa akufanelekile.
Isiphakamiso: qinisekisa ukuchaneka kokulandelelana kwe-primer kwaye i-primer ayizange ithotywe;ukuba isignali yokukhulisa i-amplification ayilungile, zama ukuthoba iqondo lokushisa le-annealing kwaye ulungelelanise i-primer concentration ngokufanelekileyo.
5.Isixa setemplate sincinci okanye sininzi kakhulu.
Isincomo: Yenza ithempleyithi yohlengahlengiso lodidi, kwaye ukhethe itemplate yoxinaniso ngeyona mpembelelo ilungileyo yePCR kulingo lwexesha lokwenyani lwePCR.
I-NTC inexabiso eliphezulu kakhulu le-fluorescence
Ungcoliseko lwe-1.Reagent olubangelwa ngexesha lokusebenza.
Isincomo: Faka indawo ngezinto ezintsha zokuvavanya ixesha lokwenyani lePCR.
2.Ungcoliseko lwenzekile ngexesha lokulungiselela inkqubo yokusabela kwe-PCR.
Isincomo: Thatha imilinganiselo yokukhusela eyimfuneko ngexesha lokusebenza, njengale: ukugqoka iiglavu ze-latex, usebenzisa i-pipetti tip kunye nesihlungi, njl.
3.Ii-primers zihlanjululwe, kwaye ukuthotywa kwee-primers kuya kubangela ukukhulisa okungangqalanga.
Ingcebiso: Sebenzisa i-SDS-PAGE i-electrophoresis ukubona ukuba iiprimers zithotyiwe, kwaye ubeke endaweni yazo ngeeprimer ezintsha zeLixesha loNyaniso lwemifuniselo yePCR.
Primer dimer okanye non-specific amplification
I-1.I-concentration ye-Mg2 + ayifanelekanga.
Isincomo: I-Mg2 + yoxinaniso lwe-2 × Real PCR Easy TM Mix esibonelela ngayo yi-3.5 mM.Nangona kunjalo, kwezinye iiprimers ezikhethekileyo kunye neetemplates, ugxininiso lweMg2 + lunokuba phezulu.Ke ngoko, unokongeza ngokuthe ngqo i-MgCl2 ukongeza i-Mg2+ yoxinaniso.Kuyacetyiswa ukuba kwandiswe i-Mg2+ 0.5mM ixesha ngalinye ukwenzela ukuba kuphuculwe.
2.Iqondo lobushushu le-PCR liphantsi kakhulu.
Ingcebiso: Yongeza iqondo lobushushu le-PCR nge-1℃ okanye nge-2℃ ngexesha ngalinye.
3.Imveliso yePCR inde kakhulu.
Isincomo: Ubude be-Real Time PCR imveliso kufuneka ibe phakathi kwe-100-150bp, ingabi ngaphezu kwe-500bp.
4.Ii-primers zihlanjululwe, kwaye ukuchithwa kwee-primers kuya kukhokelela ekubonakaleni kwe-amplification ethile.
Ingcebiso: Sebenzisa i-SDS-PAGE i-electrophoresis ukubona ukuba iiprimers zithotyiwe, kwaye ubeke endaweni yazo ngeeprimer ezintsha zeLixesha loNyaniso lwemifuniselo yePCR.
I-5.Inkqubo ye-PCR ayifanelekanga, okanye inkqubo incinci kakhulu.
Ingcebiso: Inkqubo yokusabela ye-PCR incinci kakhulu iya kubangela ukuchaneka kobhaqo ukuba kwehle.Kungcono ukusebenzisa inkqubo yokusabela ekhuthazwa sisixhobo sobungakanani bePCR ukuphinda usebenzise umfuniselo weXesha lokwenyani wePCR.
Ukuphindaphinda okulambathayo kwamaxabiso obungakanani
1.Isixhobo asisebenzi kakuhle.
Isiphakamiso: Kunokubakho iimpazamo phakathi komngxuma ngamnye we-PCR wesixhobo, okubangela ukuveliswa kakubi ngexesha lokulawula ubushushu okanye ukufumanisa.Nceda ujonge ngokwemiyalelo yesixhobo esihambelanayo.
2.Ubunyulu besampula ayilungile.
Isincomo: Iisampulu ezingcolileyo ziya kukhokelela ekuveliseni okungahambi kakuhle kovavanyo, okubandakanya ukucoceka kwetemplate kunye neeprimers.Kungcono ukuhlambulula itemplate, kwaye i-primers ihlanjululwe kakuhle yi-SDS-PAGE.
I-3.I-PCR yokulungiselela inkqubo kunye nexesha lokugcinwa lide kakhulu.
Ingcebiso: Sebenzisa inkqubo ye-PCR yeXesha lokwenyani yovavanyo lwe-PCR ngoko nangoko emva kolungiselelo, kwaye ungayishiyi ecaleni ixesha elide.
4.Iimeko zokukhulisa i-PCR azifanelekanga, kwaye ukulandelelana kwe-primer okanye ukugxininiswa akufanelekile.
Isiphakamiso: qinisekisa ukuchaneka kokulandelelana kwe-primer kwaye i-primer ayizange ithotywe;ukuba isignali yokukhulisa i-amplification ayilungile, zama ukuthoba iqondo lokushisa le-annealing kwaye ulungelelanise i-primer concentration ngokufanelekileyo.
I-5.Inkqubo ye-PCR ayifanelekanga, okanye inkqubo incinci kakhulu.
Ingcebiso: Inkqubo yokusabela ye-PCR incinci kakhulu iya kubangela ukuchaneka kobhaqo ukuba kwehle.Kungcono ukusebenzisa inkqubo yokusabela ekhuthazwa sisixhobo sobungakanani bePCR ukuphinda usebenzise umfuniselo weXesha lokwenyani wePCR.