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Foreasy HS Taq DNA Polymerase

Foreasy HS Taq DNA Polymerase Umfanekiso Featured
  • Foreasy HS Taq DNA Polymerase

Inkcazelo yeKit:

◮Ukucaciswa okuphezulu: I-enzyme enomsebenzi ophezulu wokutshisa ukuqala.

◮Ukwandiswa ngokukhawuleza: 10 sec/kb.

◮Ukuguquguquka kwetemplate ephezulu : inokusetyenziselwa ngokufanelekileyo ukukhulisa PhezuluGCixabisokwayeezahlukeneyo nzima-to-ukukhulisa DNA template.

◮Ukunyaniseka okuqinileyo: Ukunyaniseka ngamaxesha angama-6of eqhelekileyo Taq Enzyme.

amandla angaphambili


Iinkcukacha zeMveliso

Iithegi zeMveliso

FAQ

Inkcazo

I-Foreasy HS Taq DNA Polymerase yi-enzayim entsha ye-Taq echazwe kwi-Escherichia coli iibhaktheriya zobunjineli ngeteknoloji yokuhlanganiswa kwakhona kofuzo.Emva kokuba i-enzyme iphathwe ngenkqubo ekhethekileyo, yona's umsebenzi uyanqandwa phambi kokusebenza kwe-thermal, ngaloo ndlela inqanda ukukhulisa okungangcaciswanga okubangelwa yi-annealing engabonakaliyo ye-primers okanye i-primer dimers phantsi kweemeko zokushisa eziphantsi.Le mveliso ifanelekile kwi-PCR React ethileion, M ultiple x PCR , umxholo ophezulu weGC ( > 60%) ,ngeisakhiwo sesibiniokanye enyegenom yangasemva eyomeleleyoicsi-amplification kunye ne-genom enkuluicsukubhaqwa kokukhulisa.I-enzyme inomsebenzi we-5' → 3' we-DNA polymerase kunye ne-5' → 3' umsebenzi we-exonuclease, kodwa akukho 3' → 5' exonuclease.

Amacandelo ekhithi

Icandelo I-IM-01021 I-IM-01022 I-IM-01023
I-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  250 ml × 5  500 ml × 25

Iimpawu&izinto eziluncedo

- Ukucaciswa okuphezulu: I-enzyme enomsebenzi ophezulu wokutshisa ukuqala.

- Ukwandisa ngokukhawuleza: 10 sec / kb.

-Ukuguquguquka kwetemplate ephezulu: ingasetyenziselwa ukukhulisa ngokufanelekileyo PhezuluGCixabisokwayeezahlukeneyo nzima-to-ukukhulisa DNA template.

- Ukunyaniseka okuqinileyo: Ukunyaniseka ngamaxesha angama-6of eqhelekileyo Taq Enzyme.

Isicelo seKit

-Iinkqubo ezahlukeneyo zePCR/qPCR kunye nenkqubo yePCR ngqo

-Iqhekeza leDNA le-PCR

- DNA uphawu

- DNA Ulandelelwano

-PCR kunye nomsila

Inkcazo yomsebenzi

I-1U: Ubungakanani be-enzyme efunekayo ukubandakanya i-10 nmol yeDNAkwi-asidi-izinto ezinganyibilikiyo usebenzisa i-salmon esebenzayo idlozi DNA njenge template / primer, 74 °C, imizuzu engama-30.

Imeko yokusabela

Ubushushu Ixesha lokuphendula Ixesha lomjikelo
37°C 5 imiz 1
94°C 5 imiz 1
94°C 10 Imizuzwana  40
60°C 10 Imizuzwana

Phawula:Kwiinkqubo ze-10 µL kunye ne-20 µL, yongeza umthamo olinganayo we-oyile yeminerali ukuba i-thermal cycler ayinaso isiciko sobushushu.

Iimeko zokusabela kwe-PCR ziyahluka ngokuxhomekeke kwiimeko zesakhiwo setemplates, iiprimers, kunye nokunye.Kumsebenzi othile, kuyimfuneko ukuyila iimeko ezifanelekileyo zokusabela, kubandakanywa ukushisa kwe-annealing, ixesha elongezelelweyo, njl., ngokwemiqathango ethile efana nohlobo lwetemplate, ubukhulu beqhekeza ekujoliswe kulo, ukulandelelana kwesiseko seqhekeza elikhulisiwe, kunye nomxholo weGC kunye nobude be-primer.

Ugcino

-20 ± 5 °C iminyaka emi-2 okanye kwi -80 °C yokugcina ixesha elide.

  • Ngaphambili:
  • Okulandelayo:

    • Akukho miqondiso yokukhulisa

    I-1.I-Taq DNA Polymerase kwikiti ilahlekelwa ngumsebenzi wayo ngenxa yokugcinwa okungafanelekanga okanye ukuphelelwa yisikhathi kwekiti.
    Ingcebiso: Qinisekisa imiqathango yokugcina ikhithi;yongeza kwakhona imali efanelekileyo ye-Taq DNA Polymerase kwinkqubo ye-PCR okanye uthenge i-Real Time PCR Kit kwimifuniselo ehambelanayo.

    2.Zininzi ii-inhibitors ze-Taq DNA Polymerase kwi-template ye-DNA.
    Ingcebiso: Hlaziya itemplate okanye unciphise inani letemplate esetyenzisiweyo.

    3.I-concentration ye-Mg2 + ayifanelekanga.
    Isincomo: I-Mg2 + yoxinaniso lwe-2× Real PCR Mix esibonelela ngayo yi-3.5mM.Nangona kunjalo, kwezinye iiprimers ezikhethekileyo kunye neetemplates, ugxininiso lweMg2 + lunokuba phezulu.Ke ngoko, unokongeza ngokuthe ngqo i-MgCl2 ukongeza i-Mg2+ yoxinaniso.Kuyacetyiswa ukuba kwandiswe i-Mg2+ 0.5mM ixesha ngalinye ukwenzela ukuba kuphuculwe.

    4.Iimeko zokukhulisa i-PCR azifanelekanga, kwaye ukulandelelana kwe-primer okanye ukugxininiswa akufanelekile.
    Isiphakamiso: qinisekisa ukuchaneka kokulandelelana kwe-primer kwaye i-primer ayizange ithotywe;ukuba isignali yokukhulisa i-amplification ayilungile, zama ukuthoba iqondo lokushisa le-annealing kwaye ulungelelanise i-primer concentration ngokufanelekileyo.

    5.Isixa setemplate sincinci okanye sininzi kakhulu.
    Isincomo: Yenza ithempleyithi yohlengahlengiso lodidi, kwaye ukhethe itemplate yoxinaniso ngeyona mpembelelo ilungileyo yePCR kulingo lwexesha lokwenyani lwePCR.

    I-NTC inexabiso eliphezulu kakhulu le-fluorescence

    Ungcoliseko lwe-1.Reagent olubangelwa ngexesha lokusebenza.
    Isincomo: Faka indawo ngezinto ezintsha zokuvavanya ixesha lokwenyani lePCR.

    2.Ungcoliseko lwenzekile ngexesha lokulungiselela inkqubo yokusabela kwe-PCR.
    Isincomo: Thatha imilinganiselo yokukhusela eyimfuneko ngexesha lokusebenza, njengale: ukugqoka iiglavu ze-latex, usebenzisa i-pipetti tip kunye nesihlungi, njl.

    3.Ii-primers zihlanjululwe, kwaye ukuthotywa kwee-primers kuya kubangela ukukhulisa okungangqalanga.
    Ingcebiso: Sebenzisa i-SDS-PAGE i-electrophoresis ukubona ukuba iiprimers zithotyiwe, kwaye ubeke endaweni yazo ngeeprimer ezintsha zeLixesha loNyaniso lwemifuniselo yePCR.

    Primer dimer okanye non-specific amplification

    I-1.I-concentration ye-Mg2 + ayifanelekanga.
    Isincomo: I-Mg2 + yoxinaniso lwe-2 × Real PCR Easy TM Mix esibonelela ngayo yi-3.5 mM.Nangona kunjalo, kwezinye iiprimers ezikhethekileyo kunye neetemplates, ugxininiso lweMg2 + lunokuba phezulu.Ke ngoko, unokongeza ngokuthe ngqo i-MgCl2 ukongeza i-Mg2+ yoxinaniso.Kuyacetyiswa ukuba kwandiswe i-Mg2+ 0.5mM ixesha ngalinye ukwenzela ukuba kuphuculwe.

    2.Iqondo lobushushu le-PCR liphantsi kakhulu.
    Ingcebiso: Yongeza iqondo lobushushu le-PCR nge-1℃ okanye nge-2℃ ngexesha ngalinye.

    3.Imveliso yePCR inde kakhulu.
    Isincomo: Ubude be-Real Time PCR imveliso kufuneka ibe phakathi kwe-100-150bp, ingabi ngaphezu kwe-500bp.

    4.Ii-primers zihlanjululwe, kwaye ukuchithwa kwee-primers kuya kukhokelela ekubonakaleni kwe-amplification ethile.
    Ingcebiso: Sebenzisa i-SDS-PAGE i-electrophoresis ukubona ukuba iiprimers zithotyiwe, kwaye ubeke endaweni yazo ngeeprimer ezintsha zeLixesha loNyaniso lwemifuniselo yePCR.

    I-5.Inkqubo ye-PCR ayifanelekanga, okanye inkqubo incinci kakhulu.
    Ingcebiso: Inkqubo yokusabela ye-PCR incinci kakhulu iya kubangela ukuchaneka kobhaqo ukuba kwehle.Kungcono ukusebenzisa inkqubo yokusabela ekhuthazwa sisixhobo sobungakanani bePCR ukuphinda usebenzise umfuniselo weXesha lokwenyani wePCR.

    Ukuphindaphinda okulambathayo kwamaxabiso obungakanani

    1.Isixhobo asisebenzi kakuhle.
    Isiphakamiso: Kunokubakho iimpazamo phakathi komngxuma ngamnye we-PCR wesixhobo, okubangela ukuveliswa kakubi ngexesha lokulawula ubushushu okanye ukufumanisa.Nceda ujonge ngokwemiyalelo yesixhobo esihambelanayo.

    2.Ubunyulu besampula ayilungile.
    Isincomo: Iisampulu ezingcolileyo ziya kukhokelela ekuveliseni okungahambi kakuhle kovavanyo, okubandakanya ukucoceka kwetemplate kunye neeprimers.Kungcono ukuhlambulula itemplate, kwaye i-primers ihlanjululwe kakuhle yi-SDS-PAGE.

    I-3.I-PCR yokulungiselela inkqubo kunye nexesha lokugcinwa lide kakhulu.
    Ingcebiso: Sebenzisa inkqubo ye-PCR yeXesha lokwenyani yovavanyo lwe-PCR ngoko nangoko emva kolungiselelo, kwaye ungayishiyi ecaleni ixesha elide.

    4.Iimeko zokukhulisa i-PCR azifanelekanga, kwaye ukulandelelana kwe-primer okanye ukugxininiswa akufanelekile.
    Isiphakamiso: qinisekisa ukuchaneka kokulandelelana kwe-primer kwaye i-primer ayizange ithotywe;ukuba isignali yokukhulisa i-amplification ayilungile, zama ukuthoba iqondo lokushisa le-annealing kwaye ulungelelanise i-primer concentration ngokufanelekileyo.

    I-5.Inkqubo ye-PCR ayifanelekanga, okanye inkqubo incinci kakhulu.
    Ingcebiso: Inkqubo yokusabela ye-PCR incinci kakhulu iya kubangela ukuchaneka kobhaqo ukuba kwehle.Kungcono ukusebenzisa inkqubo yokusabela ekhuthazwa sisixhobo sobungakanani bePCR ukuphinda usebenzise umfuniselo weXesha lokwenyani wePCR.

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