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ISeli Ngqo RT qPCR Kit—Taqman Direct Cell Lysis Cell Ready Inyathelo elinye qRT-PCR Kits Probe

Inkcazelo yeKit:

Ikati.No.DRT-01021/01022

Kwiseli ngqo RT-qPCR usebenzisa ≤ 1000,000 iiseli,

kunye nokwenza i-RT qPCR ngqo kwiiseli ngaphandle kokucocwa kwangaphambili kwe-RNA.

Iiseli ze-lysed ngokuthe ngqo ukukhulula i-RNA ye-RT-qPCR;inkqubo yonyamezelo ephezulu yenza kube yimfuneko ukuhlanjululwa kwe-RNA kunye nokusebenzisa ngokuthe ngqo i-lysates yeseli njengeetemplates ze-RNA zokusabela kwe-RT.Ukukhawuleza kwaye kulula;uvakalelo oluphezulu, ukuchaneka okuqinileyo, kunye nokuzinza okulungileyo.

◮Ilula kwaye iyasebenza: ngeTekhnoloji ye-Cell Direct RT, iisampulu ze-RNA zinokufumaneka kwimizuzu eyi-7 nje.

Imfuno yesampulu incinci, iphantsi njengeeseli ezili-10 zinokuvavanywa.

◮Ukuphumelela okuphezulu: inokubona ngokukhawuleza i-RNA kwiiseli ezikhuliswe kwii-384, 96, 24, 12, 6-well plates.

I-DNA Eraser inokususa ngokukhawuleza i-genomes ekhutshiweyo, inciphise kakhulu impembelelo kwiziphumo zovavanyo ezilandelayo.

Inkqubo ye-RT elungiselelweyo kunye ne-qPCR yenza amanyathelo amabini e-RT-PCR aguqulelwe umva asebenze ngakumbi kwaye i-PCR icace ngakumbi, kwaye ixhathisa ngakumbi kwi-RT-qPCR reaction inhibitors.


  • :
  • Iinkcukacha zeMveliso

    Iithegi zeMveliso

    FAQ

    Khuphela izixhobo

    Iinkcazelo

    Le mveliso isebenzisa inkqubo ye-lysis buffer ekhethekileyo yokukhulula ngokukhawuleza i-RNA kwiisampuli zeseli ezikhuliswe kwi-RT-qPCR ukuphendula, ukuphelisa ixesha elide kunye nenkqubo yokucoca i-RNA enzima, kunye nemizuzu ye-7 kuphela ukufumana i-template efunekayo ye-RNA, kunye ne-5 × Direct RT Mix, 2 × Direct qPCR Mix-Taqman ebonelelwa yikiti ye-quantitive ye-PC ngokukhawuleza

    I-5 × Direct RT Mix kunye ne-2 × Direct qPCR Mix-Taqman inonyamezelo oluqinileyo lwe-inhibitor kwaye inokwenza ukuguqulwa okusebenzayo kunye nokukhulisa ngokuthe ngqo usebenzisa i-lysate yesampuli ukuba ilinganiswe njenge template.I-reagent iqulethe i-Foregene Reverse Transcriptase, i-Hot D-Taq DNA Polymerase, i-dNTPs, i-MgCl2, I-Reaction Buffer, i-PCR Optimizer kunye ne-Stabilizer, enokusetyenziswa kunye ne-lysis buffer ngokukhawuleza kwaye kulula ukubona iisampulu, kwaye ineempawu zobuntununtunu obuphezulu, ukuchaneka kunye nokuzinza.

    Iinkcukacha

    200×20μl Rxns, 1000×20μl Rxns

    Amacandelo ekhithi

    QuickEasyTMIseli ngqo RT-qPCR Kit-Taqman

    Amacandelo ekhithi

    20μl qPCR Reaction System

    DRT-01021 DRT-01022  

    Phawula

    200 T 1000 T
     

     

    Inxalenye I

    Isithinteli CL 4 ml 20 ml  

     Iseli Lysis

    Foregene Protease Plus II 80 μl 400 μl
    Isithinteli ST 400 μl 1 ml × 2
     

     

     

    Inxalenye II

    DNA Eraser 80 μl 400 μl
    5×Ngqo RT uMxube * 160 μl 800 μl RT
    2× ngokuthe ngqo qPCR Mix-Taqman * 1 ml × 2 1.7 ml × 6  

    qPCR

    I-20 × i-ROX ye-Reference Dye 40 μl 200 μl
    RNase-Free ddH2O 1.7 ml 10 ml  

    Incwadi Yemiyalelo

    Iqhekeza eli-1

    Iqhekeza eli-1

     

    *:I-Cell Lysis, i-5 × Direct RT Mix, 2× Direct qPCR Mix-Taqman inokuthengwa ngokwahlukileyo

    Iimpawu&izinto eziluncedo

    Ilula kwaye iyasebenza : ngeTekhnoloji ye-Cell Direct RT, iisampuli ze-RNA zinokufumaneka kwimizuzu eyi-7 nje.

    ∎ Imfuno yesampulu incinci, ingaphantsi njengeeseli ezili-10 zinokuvavanywa.

    ■ Ukugqithisa okuphezulu: inokubona ngokukhawuleza i-RNA kwiiseli ezikhuliswe kwii-384, 96, 24, 12, 6-well plates.

    ■ I-DNA Eraser inokususa ngokukhawuleza iigenomes ezikhutshiweyo, inciphise kakhulu impembelelo kwiziphumo zovavanyo ezilandelayo.

    ■ Inkqubo elungiselelwe i-RT kunye ne-qPCR yenza amanyathelo amabini e-RT-PCR akhutshelwe umva asebenze ngakumbi kwaye i-PCR icace ngakumbi, kwaye imelane ngakumbi ne-RT-qPCR reaction inhibitors.

    Isicelo seKit

    Ububanzi besicelo: iiseli ezikhuliswe.

    - I-RNA ekhutshwe ngesampuli ye-lysis: isebenza kuphela kwi-template ye-RT-qPCR yale khithi.

    - Ikiti ingasetyenziselwa ezi njongo zilandelayo: uhlalutyo lwe-gene expression, ukuqinisekiswa kwe-siRNA-mediated gene silence effect, ukuhlolwa kweziyobisi, njl.

    Ugcino kunye nobomi beShelf

    Inxalenye I yale khithi kufuneka igcinwe ngo-4;Icandelo II kufuneka ligcinwe kwi -20 ℃.

     I-Foregene Protease Plus II kufuneka igcinwe ngo-4℃, musa ukuba ngumkhenkce ku -20℃.

     I-Reagent 2×Direct qPCR Mix-Taqman kufuneka igcinwe kwi -20ebumnyameni;ukuba isetyenziswa rhoqo, inokugcinwa ngo-4℃ kugcino lwexesha elifutshane (sebenzisa ngaphakathi kweentsuku ezili-10).


  • Ngaphambili:
  • Okulandelayo:

  • Ixesha lokwenyani imigaqo yoyilo lwe-PCR primer

    I-Primer yokuPhambili kunye ne-Primer yokuBuyisa

    Ngexesha lokwenyani iPCR, uyilo lweprimer lubaluleke kakhulu.Iiprimers zinxulumene nobucacileyo kunye nokusebenza kakuhle kokwandiswa kwePCR, kwaye zinokuyilwa ngokubhekiselele kule migaqo ilandelayo:

    • Ubude bePrimer: 18-30bp.
    • Umxholo weGC: 40-60%.
    • Ixabiso le-Tm: I-software yoyilo lwe-Primer, njenge-Primer 5, inokunika ixabiso le-Tm le-primer.Amaxabiso e-Tm ee-primers ezinyukayo nezisezantsi kufuneka zibe kufutshane kangangoko.Ifomula yokubala ye-Tm nayo ingasetyenziswa: Tm = 4 °C (G + C) + 2 °C (A + T).Xa kusenziwa i-PCR, iqondo lobushushu elingaphantsi kwexabiso le-primer Tm le-5 °C likhethwa ngokubanzi njengeqondo lobushushu lokutshisa (ukunyuka okuhambelanayo kwiqondo lobushushu lobushushu kunokunyusa ukuchana kwe-PCR reaction).
    • Iiprimers kunye neemveliso zePCR:
    1. I-design primer PCR yokukhulisa ubude bemveliso ikhethwa yi-100-150bp.
    2. Ii-primers zoyilo kwindawo yesibini yesakhiwo setemplate kufuneka igwenywe kangangoko kunokwenzeka.
    3. Gwema ukubunjwa kweziseko ezi-2 okanye ngaphezulu ezihambelanayo phakathi kwee-3′ zeziphelo ze-primers ezinyukayo kunye nezantsi.
    4. Isiseko se-Primer 3′ asikwazi ukubakho nge-3 eyongezelelweyo elandelelanayo i-G okanye i-C.
    5. Iiprimers ngokwazo azikwazi ukuba nezakhiwo ezincedisayo, ngaphandle koko kuya kwenziwa isakhiwo se-hairpin, esichaphazela ukukhulisa i-PCR.
    6. I-ATCG kufuneka isasazwe ngokulinganayo ngokulandelelana kwe-primer, kwaye isiseko se-terminal ye-3 kufuneka sigwenywe njenge-T.

    Isihlomelo1:Cell ngqoI-RT-qPCR Ikhithi icandeloIpakethi yokongeza

    1.Cell Lysis Solution

    Iseli Lysis Solution

    Amacandelo ekhithi

    (Inkqubo ye-lysis ye-24 / kakuhle)

    DRT-01011-A1

    DRT-01011-A2

    100 T

    500 T

    InxalenyeI

    Isithinteli CL

    20 ml

    100 ml

    Foregene Protease Plus II

    400 μl

    1 ml × 2

    Isithinteli ST

    1 ml × 2

    10 ml

    InxalenyeII

    DNA Eraser

    400 μl

    1 ml × 2

    2. Umxube we-RT

    RT Umxube

    Amacandelo ekhithi

    (20 μl inkqubo yokusabela)

    DRT-01011-B1

    200 T

    5 × Ngqo RT Mix

    800 μl

    RNase-Free ddH2O

    1.7 ml × 2

    3.qPCR Mix

    Umxube we-qPCR

    Amacandelo ekhithi

    (20 μl inkqubo yokusabela)

    DRT-01021-C1

    DRT-01021-C2

    200 T

    1000 T

    2 × Direct qPCR Mix-Taqman

    1 ml × 2

    1.7 ml × 6

    I-20 × i-ROX ye-Reference Dye

    40 μl

    200 μl

    RNase-Free ddH2O

    1.7 ml

    10 ml

     

    Izikhokelo zoMyalelo:

     Iseli eKhawulezayo eNgqo ngqo i-RT-qPCR Kit-Taqman

    Bhala umyalezo wakho apha kwaye uwuthumele kuthi