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Foreasy HS Taq DNA Polymerase

Foreasy HS Taq DNA Polymerase Chithunzi Chowonetsedwa
  • Foreasy HS Taq DNA Polymerase

Kufotokozera Kwa Zida:

◮Kukhazikika kwakukulu: Enzyme yokhala ndi ntchito yoyambira yotentha kwambiri.

◮Kukulitsa Mwachangu: 10 sec/kb.

◮High template adaptability : itha kugwiritsidwa ntchito kukulitsa bwino HighGCmtengondizosiyanasiyana zovuta kukulitsa DNA template.

◮Kukhulupirika Kwambiri: Kukhulupirika ndi ka 6of wamba Taq Enzyme.

mphamvu zam'tsogolo


Tsatanetsatane wa Zamalonda

Zolemba Zamalonda

FAQ

Kufotokozera

Foreasy HS Taq DNA Polymerase ndi puloteni yatsopano ya Taq yowonetsedwa mu Escherichia coli engineering bacteria pogwiritsa ntchito ukadaulo wa gene recombination.Pambuyo enzyme kuthandizidwa ndi njira yapadera, izo's ntchito imalephereka musanayambe kuyatsa matenthedwe, motero kulepheretsa kukulitsa kosagwirizana kwenikweni komwe kumachitika chifukwa cha kusakhazikika kwapadera kwa ma primers kapena ma dimers oyambira pansi pa kutentha kochepa.Izi ndizoyenera PCR React yeniyeniion, M ultiple x PCR , high GC content (> 60%),ndidongosolo lachiwirikapena zinamaziko amphamvu genomizikukulitsa ndi genom yayikuluizikuzindikira kwa amplification.Enzyme ili ndi 5' → 3' DNA polymerase ntchito ndi 5' → 3' exonuclease ntchito, koma palibe 3' → 5' exonuclease ntchito.

Zida zamagulu

Chigawo IM-01021 IM-01022 IM-01023
Foreasy HS Taq DNA Polymerase (5 U/μL)  5000 U (1 mL)  50 KU (10 mL)  500 KU (100 mL)
2 × Taq Reaction Buffer  25mL × 5  250mL × 5  500mL × 25

Mbali & ubwino

- Kukhazikika kwakukulu: Enzyme yokhala ndi ntchito yoyambira yotentha kwambiri.

- Kukulitsa Mwachangu: 10 sec/kb.

- Kusinthasintha kwakukulu kwa template: kungagwiritsidwe ntchito kukulitsa bwino KwambiriGCmtengondizosiyanasiyana zovuta kukulitsa DNA template.

- Kukhulupirika kolimba: Kukhulupirika ndi ka 6of wamba Taq Enzyme.

Kit application

- Mitundu yosiyanasiyana ya PCR/qPCR ndi makina olunjika a PCR

- PCR Amplified DNA Fragment

- DNA chizindikiro

- Kutsata kwa DNA

- PCR kuphatikiza A mchira

Tanthauzo la Ntchito

1U : Kuchuluka kwa enzyme yofunikira kuphatikiza 10 nmol waDNAmu zinthu zosasungunuka za asidi pogwiritsa ntchito DNA ya umuna wa salmon monga template / primer, 74 °C, mphindi 30.

Mkhalidwe Wochita

Kutentha Nthawi Yochitira Nthawi yozungulira
37°C 5 min 1
94°C 5 min 1
94°C 10 Mphindi  40
60°C 10 Mphindi

Zindikirani:Pamakina a 10 µL ndi 20 µL, onjezani voliyumu yofanana yamafuta amchere ngati woyendetsa matenthedwe alibe chivindikiro cha kutentha.

Zochitika za PCR zimasiyana malinga ndi momwe ma templates amapangidwira, zoyambira, ndi zina zotero.Pogwira ntchito inayake, ndikofunikira kupanga momwe mungachitire bwino, kuphatikiza kutentha kwa annealing, nthawi yowonjezera, ndi zina zambiri, molingana ndi mikhalidwe monga mtundu wa template, kukula kwachidutswa chandamale, mndandanda wagawo lachidutswa chokulitsa, komanso zomwe zili mu GC ndi kutalika kwa choyambira.

Kusungirako

-20 ± 5 °C kwa zaka 2 kapena -80 °C posungira nthawi yaitali.

  • 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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