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Igihe nyacyo PCR Byoroshyeᵀᴹ-Taqman

Igihe nyacyo PCR Byoroshyeᵀᴹ-Taqman Ikiranga Ishusho
  • Igihe nyacyo PCR Byoroshyeᵀᴹ-Taqman
  • Igihe nyacyo PCR Byoroshyeᵀᴹ-Taqman

Ibisobanuro:

◮Byoroshye - 2 × PCR Kuvanga kugirango ugabanye ikosa ryikigereranyo nigihe cyo gukora

◮Byihariye-byateguwe neza na hot-itangira Taq enzyme irashobora gukumira amplification idasanzwe hamwe na primer dimer

◮Ubukangurambaga bukabije - bushobora kumenya kopi ntoya yicyitegererezo

◮Guhindura byinshi-bihujwe nibikoresho nyabyo-byuzuye bya PCR

imbaraga


Ibicuruzwa birambuye

Ibicuruzwa

Ibibazo

Ibisobanuro

2X Real PCR YoroshyeTMKuvanga-Taqman yatanzwe nigihe nyacyo PCR ByoroshyeTM-Taqman kit ni sisitemu nshya ya premix ikoresha probe yihariye ya fluorescente ya Real Time PCR amplification reaction, ishobora kunoza cyane ibicuruzwa byihariye no kubyumva neza.ROX itangwa nk'irangi ryo kugenzura imbere.

2X PCR YukuriTMIvanga-Taqman ririmo Foregene idasanzwe-itangira Taq ADN Polymerase.Ugereranije na misemburo isanzwe ya Taq, ifite ibyiza byo gukora neza cyane, ubushobozi bwihariye bwo kongera imbaraga hamwe nigipimo gito kidahuye.Irashobora kugabanya amplification idasanzwe no kunoza ukuri kwa PCR.

Ibisobanuro

Igihe nyacyo PCR BiroroshyeTM-Taqman

Ibigize ibikoresho (sisitemu ya 20μl)

QP-01021

QP-01022

QP-01023

QP-01024

200T

500T

1000T

2000T

2 ×PCR nyayoBiroroshyeTMKuvanga-Taqman

1 ml ×2

1.7 ml ×3

1.7 ml ×6

1.7 ml ×12

20×Irangi ryerekana irangi

200 μl

0.5ml

1 ml

1 ml × 2

DNase Yubusa ddH2O

1,7 ml

1.7 ml ×2

10 ml

20 ml

Instruction

1

1

1

1

Ibiranga ibyiza

Byoroshye - 2X PCR Kuvanga kugirango ugabanye ikosa ryikigereranyo nigihe cyo gukora

■ Byihariye - byateguwe neza na hot-itangira Taq enzyme irashobora gukumira amplification idasanzwe hamwe na primer dimer

■ Ibyiyumvo bihanitse - birashobora kumenya kopi nkeya yicyitegererezo

Ver Guhindura byinshi-bihujwe nibikoresho nyabyo-byuzuye bya PCR

Porogaramu

Isesengura rya QPCR

Urujya n'uruza rw'akazi

RT PCR-Taqman
RT PCR-Taqman igishushanyo

Igishushanyo

Ububiko nubuzima bwiza

Iki gikoresho kigomba kubikwa kure yumucyo kandi kigomba kubikwa kuri -20 ℃.Niba ikoreshwa kenshi, irashobora kandi kubikwa kuri 4 ℃ mugihe gito (iminsi 10).

  • Mbere:
  • Ibikurikira:

    • Nta kimenyetso cyo kongera imbaraga

    1.Tak ADN Polymerase ya Taq mubikoresho itakaza ibikorwa byayo kubera kubika nabi cyangwa kurangira ibikoresho.
    Icyifuzo: Emeza uburyo bwo kubika ibikoresho;ongera wongere umubare ukwiye wa Taq DNA Polymerase muri sisitemu ya PCR cyangwa ugure ibikoresho bishya bya PCR Igihe cyakorewe ubushakashatsi bujyanye.

    2.Hariho byinshi bibuza Taq DNA Polymerase mubishusho bya ADN.
    Igitekerezo: Ongera usubize inyandikorugero cyangwa ugabanye urugero rwicyitegererezo cyakoreshejwe.

    3.Ibitekerezo bya Mg2 + ntibikwiye.
    Icyifuzo: Mg2 + kwibanda kuri2 × Imvange ya PCR nyayo dutanga ni 3.5mM.Ariko, kuri bimwe bidasanzwe primers hamwe na templates, kwibanda kwa Mg2 + birashobora kuba hejuru.Kubwibyo, urashobora kongeramo MgCl2 kugirango uhindure ibitekerezo bya Mg2 +.Birasabwa kongera Mg2 + 0.5mM buri gihe kugirango optimiz.

    4.Ibihe byo kongera PCR ntabwo bikwiye, kandi primer ikurikirana cyangwa kwibandaho ntibikwiye.
    Igitekerezo: kwemeza ukuri kwa primer bikurikiranye kandi primer ntabwo yateshejwe agaciro;niba ibimenyetso bya amplification atari byiza, gerageza kugabanya ubushyuhe bwa annealing hanyuma uhindure primer yibanze neza.

    5.Ubunini bwicyitegererezo ni gito cyane cyangwa byinshi.
    Icyifuzo: Kora icyitegererezo cyerekana umurongo ugereranya, hanyuma uhitemo icyitegererezo cyibanze hamwe nibyiza bya PCR kubigeragezo nyabyo PCR.

    NTC ifite agaciro ka fluorescence cyane

    1.Ubwandure bukabije buterwa mugihe cyo gukora.
    Icyifuzo: Simbuza reagent nshya kubihe nyabyo PCR igerageza.

    2.Kwanduza kwabaye mugihe cyo gutegura sisitemu ya reaction ya PCR.
    Icyifuzo: Fata ingamba zikenewe zo kurinda mugihe cyo gukora, nka: kwambara uturindantoki twa latex, ukoresheje umuyoboro wa pipette ufite akayunguruzo, nibindi.

    3.Ibibanza byangiritse, kandi gutesha agaciro primers bizatera amplification idasanzwe.
    Igitekerezo: Koresha SDS-PAGE electrophoreis kugirango umenye niba primers yangiritse, hanyuma uyisimbuze primers nshya kubushakashatsi bwa Real Time PCR.

    Primer dimer cyangwa idasanzwe idasanzwe

    1.Ibitekerezo bya Mg2 + ntibikwiye.
    Icyifuzo: Mg2 + kwibanda kuri 2 × Real PCR EasyTM ivanze dutanga ni 3.5 mM.Ariko, kuri bimwe bidasanzwe primers hamwe na templates, kwibanda kwa Mg2 + birashobora kuba hejuru.Kubwibyo, urashobora kongeramo MgCl2 kugirango uhindure ibitekerezo bya Mg2 +.Birasabwa kongera Mg2 + 0.5mM buri gihe kugirango optimiz.

    2.Ubushyuhe bwa PCR buri hasi cyane.
    Igitekerezo: Ongera ubushyuhe bwa PCR kuri 1 ℃ cyangwa 2 ℃ buri gihe.

    3.Ibicuruzwa bya PCR ni birebire cyane.
    Icyifuzo: Uburebure bwibicuruzwa nyabyo PCR bigomba kuba hagati ya 100-150bp, ntibirenze 500bp.

    4.Ibibanza byangiritse, kandi gutesha agaciro primers bizaganisha ku kugaragara kwa amplification yihariye.
    Igitekerezo: Koresha SDS-PAGE electrophoreis kugirango umenye niba primers yangiritse, hanyuma uyisimbuze primers nshya kubushakashatsi bwa Real Time PCR.

    5. Sisitemu ya PCR idakwiye, cyangwa sisitemu ni nto cyane.
    Icyifuzo: Sisitemu ya reaction ya PCR ni nto cyane bizatera kumenya neza kugabanuka.Nibyiza gukoresha sisitemu ya reaction isabwa nigikoresho cya PCR kugirango wongere ukore igeragezwa ryigihe PCR.

    Gusubiramo nabi kwindangagaciro

    1.Igikoresho kirimo gukora nabi.
    Igitekerezo: Hashobora kubaho amakosa hagati ya buri mwobo wa PCR wigikoresho, bikavamo imyororokere mibi mugihe cyo gucunga ubushyuhe cyangwa gutahura.Nyamuneka reba ukurikije amabwiriza y'igikoresho kijyanye.

    2.Icyitegererezo cyera ntabwo ari cyiza.
    Icyifuzo: Ingero zanduye zizaganisha ku myororokere mibi yubushakashatsi, burimo ubuziranenge bwicyitegererezo na primers.Nibyiza gusubiramo inyandikorugero, kandi primers zezwa neza na SDS-PAGE.

    3.Itegurwa rya sisitemu ya PCR nigihe cyo kubika ni kirekire cyane.
    Igitekerezo: Koresha sisitemu nyayo PCR kubushakashatsi bwa PCR ako kanya nyuma yo kwitegura, kandi ntugasige kuruhande igihe kirekire.

    4.Ibihe byo kongera PCR ntabwo bikwiye, kandi primer ikurikirana cyangwa kwibandaho ntibikwiye.
    Igitekerezo: kwemeza ukuri kwa primer bikurikiranye kandi primer ntabwo yateshejwe agaciro;niba ibimenyetso bya amplification atari byiza, gerageza kugabanya ubushyuhe bwa annealing hanyuma uhindure primer yibanze neza.

    5. Sisitemu ya PCR idakwiye, cyangwa sisitemu ni nto cyane.
    Icyifuzo: Sisitemu ya reaction ya PCR ni nto cyane bizatera kumenya neza kugabanuka.Nibyiza gukoresha sisitemu ya reaction isabwa nigikoresho cya PCR kugirango wongere ukore igeragezwa ryigihe PCR.

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