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I-Cell Direct RT qPCR Kit—Taqman Direct Cell Lysis Cell Ready Isinyathelo esisodwa se-qRT-PCR Kits Probe

Incazelo yekhithi:

I-Cat.No.DRT-01021/01022

Ngeseli eqondile i-RT-qPCR esebenzisa amaseli angu-≤ 1000,000,

nokwenza i-RT qPCR ngokuqondile kumaseli ngaphandle kokuhlanzwa kwangaphambili kwe-RNA.

Amaseli afakwa lysed ngokuqondile ukuze akhulule i-RNA ye-RT-qPCR;uhlelo oluphakeme lokubekezelelana lukwenza kungadingeki ukuhlanza i-RNA nokusebenzisa ngokuqondile ama-cell lysates njengezifanekiso ze-RNA zokusabela kwe-RT.Fast and elula;ukuzwela okuphezulu, ukucaciswa okuqinile, nokuzinza okuhle.

◮Ilula futhi iyasebenza: Ngobuchwepheshe be-Cell Direct RT, amasampula e-RNA angatholwa ngemizuzu engu-7 nje.

Isidingo sesampula sincane, siphansi njengamaseli ayi-10 angahlolwa.

◮ Ukusebenza okuphezulu: ingakwazi ukubona ngokushesha i-RNA kumaseli akhuliswe kumapuleti angu-384, 96, 24, 12, 6-wemithombo.

I-DNA Eraser ingasusa ngokushesha ama-genome akhululiwe, inciphise kakhulu umthelela emiphumeleni yokuhlola elandelayo.

Isistimu ye-RT ethuthukisiwe ne-qPCR yenza ukuloba okuhlanekezela okuzinyathelo ezimbili kwe-RT-PCR kusebenze kahle futhi i-PCR icace kakhulu, futhi imelane kakhulu nama-RT-qPCR reaction inhibitors.


  • :
  • Imininingwane Yomkhiqizo

    Omaka bomkhiqizo

    FAQ

    Landa Izinsiza

    Izincazelo

    Lo mkhiqizo usebenzisa isistimu ye-lysis buffer eyingqayizivele ukuze ukhulule ngokushesha i-RNA kumasampula eseli akhulisiwe ukuze kusabela i-RT-qPCR, uqede inqubo yokuhlanza i-RNA ethatha isikhathi futhi ekhandlayo, kanye nemizuzu engu-7 kuphela ukuthola ithempulethi edingekayo ye-RNA, nge-5× Direct RT Mix, 2× Direct qPCR Mix-Taqman thola imiphumela ye-qualyntitive PCR ngokushesha nangendlela efanele.

    I-5× Direct RT Mix kanye ne-2× Direct qPCR Mix-Taqman zinokubekezelela okuqinile kwe-inhibitor futhi zingenza ukuguqulwa okuphumelelayo nokukhulisa ukukhulisa okuthile kusetshenziswa i-lysate yesampula ukuze ikalwe njengesifanekiso.I-reagent iqukethe i-Foregene Reverse Transcriptase, i-Hot D-Taq DNA Polymerase, i-dNTPs, i-MgCl2, I-Reaction Buffer, i-PCR Optimizer ne-Stabilizer, engasetshenziswa ne-lysis buffer ukuthola ngokushesha futhi kalula amasampula, futhi inezici zokuzwela okuphezulu, ukucaciswa nokuzinza.

    Imininingwane

    200×20μl Rxns, 1000×20μl rxn

    Izingxenye zekhithi

    QuickEasyTMI-Cell Direct RT-qPCR Kit-Taqman

    Izingxenye zekhithi

    20μl qPCR Reaction System

    I-DRT-01021 I-DRT-01022  

    Qaphela

    200 T 1000 T
     

     

    Ingxenye I

    Isivimbeli CL 4 ml 20 ml  

     Iseli Lysis

    I-Foregene Protease Plus II awu 80l 400 μl
    Isilondolozi se-ST 400 μl 1 ml × 2
     

     

     

    Ingxenye II

    I-DNA Eraser awu 80l 400 μl
    5×Imiksi ye-RT eqondile * 160 μl 800 μl RT
    2× Direct qPCR Mix-Taqman * 1 ml × 2 1.7 ml × 6  

    qPCR

    I-20 × ROX Ireferensi Idayi 40 ml 200 μl
    I-ddH yamahhala ye-RNase2O 1.7 ml 10 ml  

    Incwadi Yeziqondiso

    1 isiqephu

    1 isiqephu

     

    *:I-Cell Lysis, 5×Direct RT Mix, 2× Direct qPCR Mix-Taqman angathengwa ngokuhlukana

    Izici&izinzuzo

    Kulula futhi kusebenza ngempumelelo : Ngobuchwepheshe be-Cell Direct RT, amasampula e-RNA angatholwa ngemizuzu engu-7 nje.

    ■ Isidingo sesampula sincane, siphansi njengoba amaseli ayi-10 angahlolwa.

    ■ Ukuphuma okuphezulu: ikwazi ukubona ngokushesha i-RNA kumaseli akhiwe kumapuleti angu-384, 96, 24, 12, 6-wemithombo.

    ■ I-DNA Eraser ingasusa ngokushesha ama-genome akhululiwe, inciphise kakhulu umthelela emiphumeleni yokuhlola elandelayo.

    ■ Isistimu ye-RT ethuthukisiwe kanye ne-qPCR yenza ukuloba okuhlanekezela okuzinyathelo ezimbili kwe-RT-PCR kusebenze kahle futhi i-PCR icace kakhulu, futhi imelane kakhulu ne-RT-qPCR reaction inhibitors.

    Isicelo sekhithi

    Ububanzi bokusebenza: amaseli akhulisiwe.

    - I-RNA ekhishwe ngesampula ye-lysis: isebenza kuphela kusifanekiso se-RT-qPCR sale kit.

    - Ikhithi ingasetshenziselwa lezi zinhloso ezilandelayo: ukuhlaziywa kwesisho sofuzo, ukuqinisekiswa komphumela wokuthulisa ufuzo we-siRNA-mediated, ukuhlolwa kwezidakamizwa, njll.

    Isitoreji kanye Nempilo yeshelufu

    Ingxenye I yale khithi kufanele igcinwe ku-4;Ingxenye II kufanele igcinwe ku -20 ℃.

     I-Foregene Protease Plus II kufanele igcinwe ku-4℃, ungafrizi ku -20℃.

     I-Reagent 2×I-Direct qPCR Mix-Taqman kufanele igcinwe ku -20ebumnyameni;uma isetshenziswa njalo, ingabuye igcinwe ku-4℃ ukuze ugcine isikhathi esifushane (sebenzisa phakathi 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:
    1. I-design primer PCR ubude bomkhiqizo wokukhulisa u-100-150bp.
    2. Iziqalo zokuklama endaweni yesibili yesakhiwo sesifanekiso kufanele zigwenywe kakhulu ngangokunokwenzeka.
    3. Gwema ukwakheka kwezisekelo ezi-2 noma ngaphezulu ezihambisanayo phakathi kwamaphethelo angu-3′ weziqalisi ezikhuphuka nomfula.
    4. Isisekelo setheminali ye-Primer 3′ asikwazi ukuba khona nezinye ezingu-3 ezengeziwe ezilandelanayo ze-G noma C.
    5. Ama-primer ngokwawo awakwazi ukuba nezakhiwo ezihambisanayo, ngaphandle kwalokho kuzokwakhiwa isakhiwo se-hairpin, esithinta ukukhuliswa kwe-PCR.
    6. I-ATCG kufanele isatshalaliswe ngokulinganayo ngangokunokwenzeka ekulandeleni kokuqala, futhi isisekelo setheminali esingu-3′ kufanele sigwenywe njengo-T.

    Isithasiselo1:Cethi DirectI-RT-qPCR Ingxenye yekhithit iphakethe le-supplement

    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. I-RT Mix

    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-01021-C1

    I-DRT-01021-C2

    200 T

    1000 T

    2× Direct qPCR Mix-Taqman

    1 ml × 2

    1.7 ml × 6

    I-20 × I-ROX Reference Dye

    40 ml

    200 μl

    I-ddH yamahhala ye-RNase2O

    1.7 ml

    10 ml

     

    Amamanuwali wemiyalo:

     I-Quick Easy Cell Direct RT-qPCR Kit-Taqman

    Bhala umyalezo wakho lapha futhi usithumelele wona