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Real Time PCR Easyᵀᴹ-Taqman

Real Time PCR Easyᵀᴹ-Taqman Chithunzi Chowonetsedwa
  • Real Time PCR Easyᵀᴹ-Taqman
  • Real Time PCR Easyᵀᴹ-Taqman

Kufotokozera Kwa Zida:

◮Zosavuta—2 × PCR Mix kuti muchepetse zolakwika zoyeserera ndi nthawi yogwira ntchito

◮Specific-optimized buffer ndi hot-start Taq enzyme imatha kuteteza kukulitsa kosakhazikika komanso mapangidwe a dimer.

◮Kumverera kwakukulu - kumatha kuzindikira makope otsika a template

◮Kusinthasintha kwabwino - kumagwirizana ndi zida zenizeni zenizeni za PCR

mphamvu zam'tsogolo


Tsatanetsatane wa Zamalonda

Zolemba Zamalonda

FAQ

Mafotokozedwe a Kit

2X Real PCR YosavutaTMMix-Taqman yoperekedwa ndi Real Time PCR EasyTM-Taqman kit ndi dongosolo latsopano la premix lomwe limagwiritsa ntchito ma probes apadera a fulorosenti pakuchita kwa Real Time PCR amplification, yomwe imatha kusintha kwambiri tsatanetsatane wazinthu komanso kukhudzidwa kwamachitidwe.ROX imaperekedwa ngati utoto wowongolera mkati.

2X Real PCR YosavutaTMMix-Taqman ili ndi Taq DNA Polymerase yapadera ya Foregene.Poyerekeza ndi ma enzyme wamba a Taq, ili ndi zabwino zake pakukweza kwakukulu, kuthekera kokulirapo kwamphamvu komanso kutsika kosagwirizana.Itha kuchepetsa kukulitsa kosakhazikika ndikuwongolera kulondola kwa PCR.

Zofotokozera

Real Time PCR EasyTM- Taqman

Kapangidwe ka zida (20μl system)

QP-01021

QP-01022

QP-01023

QP-01024

200T

500T

1000T

2000T

2 × paPCR yeniyeniZosavutaTMSakanizani- Taqman

1 ml ×2

1.7 ml × pa3

1.7 ml × pa6

1.7 ml × pa12

20×ROX Reference Dye

200 μl

0.5ml

1 ml

1 ml × 2

DNase-Free ddH2O

1.7 ml

1.7 ml × pa2

10 ml

20 ml

Imalangizo

1

1

1

1

Mbali & ubwino

■ Yosavuta—2X PCR Mix kuti muchepetse zolakwika zoyeserera ndi nthawi yogwira ntchito

■ Specific-optimized buffer ndi hot-start Taq enzyme imatha kuletsa kukulitsa kosakhazikika komanso kupangika kwa dimer.

■ Kuzindikira kwakukulu-kumatha kuzindikira makope otsika a template

■ Kusinthasintha kwabwino—kumagwirizana ndi zida zambiri zapanthawi yeniyeni za PCR

Kit application

qPCR kusanthula

Kuyenda kwa ntchito

RT PCR-Taqman
Zithunzi za RT PCR-Taqman

Zithunzi

Kusungirako ndi moyo wa alumali

Chidachi chiyenera kusungidwa kutali ndi kuwala ndipo chiyenera kusungidwa pa -20 ℃.Ngati imagwiritsidwa ntchito pafupipafupi, imathanso kusungidwa ku 4 ℃ kwakanthawi kochepa (masiku 10).

  • Zam'mbuyo:
  • Ena:

    • Palibe zizindikiro za amplification

    1.Taq DNA Polymerase mu kit imataya ntchito yake chifukwa cha kusungirako kosayenera kapena kutha kwa zida.
    Malangizo: Tsimikizirani zosungirako zida;onjezaninso kuchuluka koyenera kwa Taq DNA Polymerase kudongosolo la PCR kapena mugule Real Time PCR Kit yatsopano kuti muyesere.

    2.Pali zoletsa zambiri za Taq DNA Polymerase mu template ya DNA.
    Yesani: Yeretsaninso template kapena kuchepetsa kuchuluka kwa template yomwe yagwiritsidwa ntchito.

    3.Kuwerengera kwa Mg2 + sikoyenera.
    Malangizo: Kuphatikizika kwa Mg2+ kwa2× Real PCR Mix yomwe timapereka ndi 3.5mM.Komabe, pazoyambira zina zapadera ndi ma templates, ndende ya Mg2 + ikhoza kukhala yokwezeka.Chifukwa chake, mutha kuwonjezera mwachindunji MgCl2 kuti mukwaniritse ndende ya Mg2+.Ndibwino kuti muwonjezere Mg2+ 0.5mM nthawi iliyonse kuti muwongolere.

    4.Kukulitsa kwa PCR sikuli koyenera, ndipo kutsatizana koyambirira kapena kusakanikirana sikuli koyenera.
    Malingaliro: tsimikizirani kulondola kwa mndandanda wa zoyambira ndipo choyambira sichinawonongeke;ngati chizindikiro chokulitsa sichili bwino, yesani kuchepetsa kutentha kwa annealing ndikusintha ndende yoyambira bwino.

    5.Kuchuluka kwa template ndi kochepa kwambiri kapena kochuluka.
    Malangizo: Chitani template ya linearization gradient dilution, ndikusankha template yomwe ili ndi PCR yabwino kwambiri pakuyesa kwa Real Time PCR.

    NTC ili ndi mtengo wokwera kwambiri wa fulorosenti

    1.Reagent kuipitsidwa chifukwa pa ntchito.
    Malangizo: M'malo ndi zoyeserera zatsopano za Real Time PCR.

    2.Kuwonongeka kunachitika panthawi yokonzekera machitidwe a PCR.
    Malangizo: Tengani njira zodzitetezera panthawi yogwira ntchito, monga: kuvala magolovesi a latex, kugwiritsa ntchito nsonga ya pipette yokhala ndi fyuluta, ndi zina zotero.

    3.Zoyambira zimawonongeka, ndipo kuwonongeka kwa zoyambira kumayambitsa kukulitsa kosagwirizana.
    Malingaliro: Gwiritsani ntchito electrophoresis ya SDS-PAGE kuti muwone ngati zoyambira zawonongeka, ndikusintha ndi zoyambira zatsopano zoyeserera za Real Time PCR.

    Primer dimer kapena matalikidwe osakhala enieni

    1.Kuwerengera kwa Mg2 + sikoyenera.
    Malangizo: Mg2 + ndende ya 2 × Real PCR EasyTM Mix yomwe timapereka ndi 3.5 mM.Komabe, pazoyambira zina zapadera ndi ma templates, ndende ya Mg2 + ikhoza kukhala yokwezeka.Chifukwa chake, mutha kuwonjezera mwachindunji MgCl2 kuti mukwaniritse ndende ya Mg2+.Ndibwino kuti muwonjezere Mg2+ 0.5mM nthawi iliyonse kuti muwongolere.

    2.The PCR annealing kutentha ndi otsika kwambiri.
    Yesani: Wonjezerani kutentha kwa PCR ndi 1 ℃ kapena 2 ℃ nthawi iliyonse.

    3.The PCR mankhwala ndi yaitali kwambiri.
    Malangizo: Kutalika kwa mankhwala a Real Time PCR kuyenera kukhala pakati pa 100-150bp, osapitirira 500bp.

    4.Zoyambira zimawonongeka, ndipo kuwonongeka kwa zoyambira kudzatsogolera ku maonekedwe a kukulitsa kwapadera.
    Malingaliro: Gwiritsani ntchito electrophoresis ya SDS-PAGE kuti muwone ngati zoyambira zawonongeka, ndikusintha ndi zoyambira zatsopano zoyeserera za Real Time PCR.

    5.Dongosolo la PCR ndi losayenera, kapena dongosolo ndi laling'ono kwambiri.
    Malingaliro: Makina a PCR ndi ochepa kwambiri apangitsa kuti kuzindikira kuchepe.Ndi bwino kugwiritsa ntchito kachitidwe kachitidwe kolimbikitsidwa ndi chida chochulukira cha PCR kuti muyambitsenso kuyesa kwa Real Time PCR.

    Kusabwerezabwereza kolakwika kwa kachulukidwe

    1.Chidacho sichikuyenda bwino.
    Lingaliro: Pakhoza kukhala zolakwika pakati pa dzenje lililonse la PCR la chida, zomwe zimapangitsa kuti pakhale kusabereka bwino pakuwongolera kutentha kapena kuzindikira.Chonde fufuzani molingana ndi malangizo a chida chofananira.

    2.Chiyerekezo cha chiyero sichili chabwino.
    Langizo: Zitsanzo zodetsedwa zipangitsa kuti kuyesako kusakhalenso bwino, komwe kumaphatikizapo chiyero cha template ndi zoyambira.Ndikwabwino kuyeretsanso template, ndipo zoyambira zimatsukidwa bwino ndi SDS-PAGE.

    3.Kukonzekera kwadongosolo la PCR ndi nthawi yosungira ndi yaitali kwambiri.
    Yesani: Gwiritsani ntchito makina a Real Time PCR poyesera PCR mukangokonzekera, ndipo musayisiye pambali kwa nthawi yayitali.

    4.Kukulitsa kwa PCR sikuli koyenera, ndipo kutsatizana koyambirira kapena kusakanikirana sikuli koyenera.
    Malingaliro: tsimikizirani kulondola kwa mndandanda wa zoyambira ndipo choyambira sichinawonongeke;ngati chizindikiro chokulitsa sichili bwino, yesani kuchepetsa kutentha kwa annealing ndikusintha ndende yoyambira bwino.

    5.Dongosolo la PCR ndi losayenera, kapena dongosolo ndi laling'ono kwambiri.
    Malingaliro: Makina a PCR ndi ochepa kwambiri apangitsa kuti kuzindikira kuchepe.Ndi bwino kugwiritsa ntchito kachitidwe kachitidwe kolimbikitsidwa ndi chida chochulukira cha PCR kuti muyambitsenso kuyesa kwa Real Time PCR.

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