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Ixesha lokwenyani PCR Easyᵀᴹ-Taqman

Ixesha lokwenyani PCR Easyᵀᴹ-Taqman Featured Image
  • Ixesha lokwenyani PCR Easyᵀᴹ-Taqman
  • Ixesha lokwenyani PCR Easyᵀᴹ-Taqman

Inkcazelo yeKit:

◮Elula—2× 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-luyakwazi ukubona iikopi eziphantsi zetemplate

◮Ukuguquguquka okuhle-kuyahambelana nezixhobo ezininzi zePCR zexesha lokwenyani

amandla angaphambili


Iinkcukacha zeMveliso

Iithegi zeMveliso

FAQ

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

2×I-PCR yokwenyaniKululaTMXuba-Taqman

1 ml ×2

1.7 ml ×3

1.7 ml ×6

1.7 ml ×12

20×I-ROX Reference Daye

200 μl

0.5ml

1 ml

1 ml × 2

DNase-Free ddH2O

1.7 ml

1.7 ml ×2

10 ml

20 ml

Iumyalelo

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

RT PCR-Taqman
Umzobo we-RT PCR-Taqman

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).

  • Ngaphambili:
  • Okulandelayo:

    • 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.

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