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QuickEasyᵀᴹ Real Time PCR Kit-Taqman

QuickEasyᵀᴹ Real Time PCR Kit-Taqman Isithombe Esifakiwe
  • QuickEasyᵀᴹ Real Time PCR Kit-Taqman

Incazelo yekhithi:

◮Isistimu yokwenza kahle ye-PCR yenza i-2×QuickEasyTMI-Real PCR Mix-Taqman ihambisana kakhulu.

◮I-Hot-start Foregene HS Taq Polymerase inekhono eliphezulu lokukhulisa i-amplification, ukuzwela kwe-amplification ephakeme, kanye nokucaciswa kokukhulisa okuphezulu.

◮2 × QuickEasyTMI-Real PCR Mix -Taqman isebenzisa isistimu eyingqayizivele elungiselelwe i-Foregene ukuze kuthuthukiswe ukuzwela kanye nokucaciswa kokutholwa kophenyo okulandelanayo, olungasetshenziselwa ukunqunywa kwe-genotyping nokukopisha ukuhluka kwenombolo nokuthola imiphumela enembayo.

◮Lo mkhiqizo uza nodayi wereferensi wangaphakathi we-ROX, ongasetshenziswa ukuqeda ingemuva lesignali kanye nephutha lesiginali phakathi kwemithombo, okulula ukuthi amakhasimende ayisebenzise ezinhlotsheni ezahlukene zamathuluzi e-PCR amaningi.

amandla angaphambili


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Imininingwane Yomkhiqizo

Omaka bomkhiqizo

FAQ

Izincazelo

I-2× QuickEasyTMI-Real PCR Mix-Taqman ihlinzekwa yi-QuickEasyTMIkhithi ye-PCR ye-Real Time ye-PCR Kit-Taqman iwuhlelo olusha lwesizukulwane oluyinhlanganisela oluklanyelwe ukusabela kokukhulisa i-Real Time PCR kusetshenziswa ama-fluorescent probes athile.Isisekelo sayo se-Foregene HS Taq DNA Polymerase sisekelwe ekuguqulweni kwe-Antibody, esetshenziswa kanye ne-Foregene ethuthukisiwe nohlelo lwe-PCR, inokucaciswa okuphezulu nokusebenza kahle kokukhulisa.Uma iqhathaniswa nengxube evamile ye-PCR, inamandla okukhulisa amandla aqinile nezinga lokungafani eliphansi.Ingasetshenziswa ku-fluorescence quantitative PCR reaction ukuze kwehliswe ukukhulisa okungaqondile nokuthuthukisa ukunemba kwe-PCR.Ingathuthukisa kakhulu ukucaciswa komkhiqizo nokuzwela kokusabela.Ngasikhathi sinye, i-ROX yanikezwa njengodayi wereferensi wangaphakathi.

I-QuickEasyTMIkhithi ye-PCR ye-Real Time ye-Kit-Taqman inobubanzi obubanzi bokusebenza enanini lesifanekiso, futhi ingathola ijika elijwayelekile elikahle ezindaweni ezihlukene zobuningi, futhi ingakwazi ukulinganisa ngokunembile futhi ithole izakhi zofuzo eziqondiwe ngokuphindaphinda okuhle nokuzethemba okuphezulu.

Imininingwane

200 Prep,500 Prep,1000 Preps,2000 Preps

Izingxenye zekhithi

QuickEasyTMIsikhathi Sangempela PCR Kit-Taqman(20μluhlelo)

Okuqukethwe kwekhithi

QP-01121

QP-01122

QP-01123

QP-01124

200 Preps

500 Preps

1000 Preps

2000 Preps

2×NgokusheshaKululaTM I-PCR yangempela Hlanganisa-Taqaman

1 mL×2

1.7 mL×3

1.7 mL×6

1.7 mL×12

20×I-ROX Reference Dye

200μL

0.5 ml

1 mL

1 mL×2

I-DNase-Free ddH2O

1.7 mL

1.7 mL×2

10 ml

20 ml

IFU

1ucezu

1ucezu

1ucezu

1ucezu

Izici&izinzuzo

■ Uhlelo oluyingqayizivele lwe-PCR lwenza i-2×QuickEasyTMI-Real PCR Mix-Taqman ihambisana kakhulu.

■ I-Hot-start I-Foregene HS Taq Polymerase inekhono eliphezulu lokukhulisa i-amplification, ukuzwela kokukhulisa ukuphakama, kanye nokucaciswa kokukhulisa ukuphakama okuphezulu.

■ 2× QuickEasyTM I-Real PCR Mix -Taqman isebenzisa isistimu eyingqayizivele elungiselelwe i-Foregene ukuze kuthuthukiswe ukuzwela kanye nokucaciswa kokutholwa kophenyo okulandelanayo, olungasetshenziselwa ukunqunywa kwe-genotyping nokukopisha ukuhluka kwenombolo nokuthola imiphumela enembayo.

■ Lo mkhiqizo uza nodayi wereferensi wangaphakathi we-ROX, ongasetshenziswa ukuqeda ingemuva lesignali kanye nephutha lesignali phakathi kwemithombo, okulula ukuthi amakhasimende ayisebenzise ezinhlotsheni ezahlukene zamathuluzi e-PCR amaningi.

Isicelo sekhithi

■ I-Fluorescence quantitative PCR yokuhlaziya ubuningi besifanekiso;

■ Cukukhuliswa kwe-PCR okuzenzakalelayo;

■ Ingasetshenziselwa ukutholwa kwe-allele.

Isitoreji kanye Nempilo yeshelufu

Ikhithi kufanele igcinwe ku -20 ° C ebumnyameni;uma isetshenziswa njalo, ingabuye igcinwe ku-4°C isikhathi esifushane (isetshenziswe ngaphakathi kwezinsuku eziyi-10).

  • Okwedlule:
  • Olandelayo:

    • Izimiso ze-Real Time PCR primer design

    Dlulisela phambili i-Primer kanye ne-Primer Reverse

    Nge-Real Time PCR, ukuklama kokuqala kubaluleke kakhulu.Ama-Primers ahlobene nokucaciswa nokusebenza kahle kokukhulisa i-PCR, futhi angaklanywa ngokubhekiselwa kule migomo elandelayo:

    Ubude be-Primer: 18-30bp.

    Okuqukethwe kwe-GC: 40-60%.

    Inani le-Tm: Isofthiwe yokuklama i-Primer, njenge-Primer 5, inganikeza inani le-Tm le-primer.Amanani e-Tm okuqalisa akhuphuka nomfula kufanele abe seduze ngangokunokwenzeka.Ifomula yokubala ye-Tm nayo ingasetshenziswa: Tm = 4 °C (G + C) + 2 °C (A + T).Lapho kwenziwa i-PCR, izinga lokushisa elingaphansi kwenani lokuqala le-Tm elingu-5 °C livame ukukhethwa njengezinga lokushisa lokukhipha isisu (ukwenyuka okuhambisanayo kwezinga lokushisa le-anneal kungakhuphula ukucaciswa kokusabela kwe-PCR).

    Ama-Primers kanye nemikhiqizo ye-PCR:

    I-design primer PCR ubude bomkhiqizo wokukhulisa u-100-150bp.

    Iziqalo zokuklama endaweni yesibili yesakhiwo sesifanekiso kufanele zigwenywe kakhulu ngangokunokwenzeka.

    Gwema ukwakheka kwezisekelo ezi-2 noma ngaphezulu ezihambisanayo phakathi kwamaphethelo angu-3′ weziqalisi ezikhuphuka nomfula.

    Isisekelo setheminali ye-Primer 3′ asikwazi ukuba khona nezinye ezingu-3 ezengeziwe ezilandelanayo ze-G noma C.

    Ama-primer ngokwawo awakwazi ukuba nezakhiwo ezihambisanayo, ngaphandle kwalokho kuzokwakhiwa isakhiwo se-hairpin, esithinta ukukhuliswa kwe-PCR.

    I-ATCG kufanele isatshalaliswe ngokulinganayo ngangokunokwenzeka ekulandeleni kokuqala, futhi isisekelo setheminali esingu-3′ kufanele sigwenywe njengo-T.

    Isithasiselo 1:Iphakethe lengxenye ye-Cell Direct RT-qPCR Kit

    1.Cell Lysis Solution


    Cell Lysis Solution

    Izingxenye zekhithi

    (Isistimu ye-lysis ye-24 / kahle)

    I-DRT-01011-A1

    I-DRT-01011-A2

    100 T

    500 T

    IngxenyeI

    Isivimbeli CL

    20 ml

    100 ml

    I-Foregene Protease Plus II

    400 μl

    1 ml × 2

    Isilondolozi se-ST

    1 ml × 2

    10 ml

    IngxenyeII

    I-DNA Eraser

    400 μl

    1 ml × 2

    2.RT Imiksi


    I-RT Mix

    Izingxenye zekhithi

    (Isistimu yokusabela engu-20 μl)

    I-DRT-01011-B1

    200 T

    5× Imiksi ye-RT eqondile

    800 μl

    I-ddH yamahhala ye-RNase2O

    1.7 ml × 2

     

    3.qPCR Mix


    I-qPCR Mix

    Izingxenye zekhithi

    (Isistimu yokusabela engu-20 μl)

    I-DRT-01011-C1

    I-DRT-01011-C2

    200 T

    1000 T

    2× Direct qPCR Mix-SYBR

    1 ml × 2

    1.7 ml × 6

    I-50 × I-ROX Reference Dye

    40 ml

    200 μl

    I-ddH yamahhala ye-RNase2O

    1.7 ml

    10 ml
    Bhala umyalezo wakho lapha futhi usithumelele wona

    OkuhlobeneImikhiqizo

    Chofoza u-enter ukuze useshe noma u-ESC ukuze uvale