Ultra HiFidelity PCR Kit

Botšepehi bo phahameng, boits'oaro bo phahameng le ts'ebetso e phahameng ea ho qala PCR ea pele.

Ultra HiFidelity PCR Kit ke sehlomathiso se secha sa kholiseho e phahameng ea PCR se loketseng khokahano le phumano e amanang le PCR. Ultra HiFi DNA Polymerase e fumanehang ka har'a kit ke DNA polymerase e ncha e potlakileng le e phahameng e entsoeng ke theknoloji e laoloang ea kholo ea limolek'hule. E ntlafatsa kamano ea DNA polymerase ho li-template, e ntlafatsa lebelo la ho matlafatsa le bokhoni ba ho atolosa enzyme, mme e eketsa sekhahla sa katleho ea PCR le lihlahisoa tsa lihlahisoa.

Katse. Che	Paka Size
4992970	1ml
4992971	5 * 1ml
4992978	5 * 5 * 1ml

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Product Qaqileng

Mohlala oa Teko

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Lihlahisoa tsa Product

Likaroloana

■ Ho sebetsa bonolo
■ Botšepehi bo phahameng: Botšepehi ke makhetlo a 50 ho feta a Taq polymerase.
■ High tobileng: Excellent tshebetso hot-simolleng ho etsa bonnete ba sehlahisoa itseng.
■ Kholiso e potlakileng: Lebelo la katoloso le ka fihla ho 10-15 sec / kb.
■ Ho atolosoa ka matla: Ho ka eketsoa likhechana tsa DNA tse ka bang 20.
■ Ts'ebetsong e pharalletseng: Setsi sena se na le PCR Enhancer mme se loketse kholiso ea li-GC tse phahameng le litempele tse rarahaneng.

Ga tlhaloso

Mofuta: Botšepehi bo phahameng ba DNA Polymerase
Lebelo la ho eketsa: 10-15 sec / kb
Boholo ba sekhechana: <20kb
Likopo: Kholiseho e phahameng ea PCR, kholiso ea liphatsa tsa lefutso, kholiso e phahameng ea GC, liphatsa tsa lefutso tsa li-genomes tse rarahaneng, cDNA e phahameng ea bots'epehi, ho bonoa ha SNP, phetoho e ikhethang ea sebaka, jj.
Tlhahiso ea DNA ho Mefuta e fapaneng ea Limela:
Tlhokomeliso: chai ea DNA e ipapisitse le mefuta ea mehlala. Lisebelisoa tsohle tse kaholimo li tsoa makhasi a bonolo.

Lihlahisoa tsohle li ka fetoloa bakeng sa ODM / OEM. Bakeng sa lintlha,ka kopo tlanya Customized Service (ODM / OEM)

E fetileng: 2 × Pfu PCR Motsoako

E 'ngoe: Sesebelisoa sa QuantScript RT

	Ho qala ka cheseho ho netefatsa sehlahisoa se ikhethang Setšoantšo sa 1. Ultra HiFi e na le ts'ebetso e ntle ea ho qala ho chesa ho etsa bonnete ba sehlahisoa sa sehlahisoa Mokhoa oa li-beacon tsa limolek'hule o sebelisitsoe (Ma et al., Anal Biochem, 2006).
	Botšepehi bo phahameng haholo, makhetlo a 50 a phahame ho feta Taq Polymerase Setšoantšo sa 2. Tšepahalo ea Ultra HiFi e phahame makhetlo a 50 ho feta ea Taq polymerase e tloaelehileng. Botšepehi ba polymerization ea Taq polymerase (ntle le ts'ebetso ea ho e lokisa) e sebelisoa e le ts'upiso.
	Kholiso e potlakileng le likhechana tse telele li ka holisoa kapele Setšoantšo sa 3. Hi HiFi e ka atoloha ho fihla ho sec 5 / kb bakeng sa likhechana tse nyane ho feta 4 kb. Bakeng sa likhechana tse telele, nako ea kholiso e ka atolosoa ka nepo. Bakeng sa likhechana tse fetang 15 kb, lebelo leo e ka bang 30 sec / kb. M: TIANGEN D15000 Lesupa
	Bokahohle bo matla le boits'oaro bo phahameng, ho bonolo ho li bala GC le likhechana tse telele tse tsoang mehloling e fapaneng Setšoantšo sa 4. Liphetho tsa kholiso ea Ultra HiFi Liphetho tsa kholiso ea enzyme ea Hi-Fi ea Morekisi K Liphetho tsa kholiso ea enzyme ea Hi-Fi ea Mofani oa thepa N M: TIANGEN D15000 Lesupa Tsela ea 1-5. Liphello tsa ntlafatso ea li-template tse bolelele bo fapaneng: 1. 750 bp; 2. 1 kb; 3. 2 kb; 4. 4 kb; 5. 6 kb Lane 6. Phello ea tlatsetso ea template e phahameng ea GC: 1915 bp (GC%: 70%); Tsela ea 7-11. Phello ea ntlafatso ea li-template tsa 2 kb tse tsoang ho genome e fapaneng: 7. Rat; 8. Raese; 9. Koro; 10. Poone; 11. Libaktheria; Tsela ea 12-14. 8 kb sephetho sa ho holisa sekhechana se telele: 12. Raese; 13. Poone;

Q: Ha ho na lihlopha tsa matlafatso

Setšoantšo sa A-1

■ Template e na le litšila tsa protheine kapa li-inhibitors tsa Taq, jj. ——Tlhoekisa template ea DNA, tlosa litšila tsa protheine kapa u ntše template ea DNA ka lisebelisoa tsa tlhoekiso.

■ Tlatsetso ea template ha e felle ——Ho eketsa ka mokhoa o nepahetseng mocheso oa "denaturation" le ho lelefatsa nako ea denaturation.

■ Ho senyeha ha litempele ——Reka template hape.

Tlhahiso ea A-2

■ Boleng bo tlase ba li-primers ——Re kopanya hape selelekela.

■ Ho senyeha ha li-primer——Fana ka litlhahlobo tse phahameng tsa likhakanyo hore e be molumo o monyane bakeng sa ho li boloka. Qoba ho hatsela hangata le ho qhibiliha kapa nako e telele ea 4 ° C e bolokiloeng.

■ Moralo o sa nepahalang oa litlhahlobo (mohlala, bolelele ba lithuso ha boa lekana, botenya bo thehiloe lipakeng tsa lintho tsa pele, jj.

EA-3 Mg²⁺ho tsepamisa maikutlo

■ Mg²⁺ ho tsepamisa mohopolo ho tlase haholo ——Ho eketsa hantle Mg²⁺mahloriso: Lokisetsa Mg²⁺ mahloriso ka letoto la likarabelo ho tloha ho 1 mM ho isa ho 3 mM ka karohano ea 0.5 mM ho fumana Mg²⁺ mahloriso bakeng sa template le primer ka 'ngoe.

Mocheso oa ho koala oa A-4

■ Mocheso o phahameng oa ho koala o ama tlamo ea selelekela le template. Fokotsa mocheso o kenang 'me u ntlafatse boemo ka gradient ea 2 ° C.

Nako ea katoloso ea A-5

■ Nako e khuts'oane ea katoloso —- Eketsa nako ea katoloso.

P: Ho ba le tšoaetso ea bohata

Phenomena: Mehlala e mebe e bonts'a lihlopha tsa tatellano.

Tšilafalo ea A-1 ea PCR

■ Tšilafalo ea sefapano ea tatellano ea sepheo kapa lihlahisoa tsa ntlafatso ——Tlhokomelo hore o se ke oa tšela sampole e nang le tatellano ea sepheo sa sampuli e fosahetseng kapa oa e tšolla ka har'a tube ea centrifuge. Li-reagents kapa lisebelisoa li lokela ho etsoa mochini o ikemetseng ho felisa li-acid tsa nucleic, mme boteng ba tšilafalo bo lokela ho khethoa ka liteko tse mpe tsa taolo.

■ Tšilafalo ea li-reagent ——Qobella li-reagents ebe u li boloka ka mocheso o tlase.

A-2 Tona-khōlōr

■ Mg²⁺ ho tsepamisa mohopolo ho tlase haholo ——Ho eketsa hantle Mg²⁺ mahloriso: Lokisetsa Mg²⁺ mahloriso ka letoto la likarabelo ho tloha ho 1 mM ho isa ho 3 mM ka karohano ea 0.5 mM ho fumana Mg²⁺ mahloriso bakeng sa template le primer ka 'ngoe.

■ Moralo oa semmuso o sa nepahalang, 'me tatellano ea sepheo e na le thuto ea bongoli le tatellano e sa amaneng le sepheo. ——Litlhahlobo tsa pele-pele.

Q: Kholiso e sa tobang

Phenomena: Lihlopha tsa kholiso ea PCR ha li lumellane le boholo bo lebelletsoeng, ebang ke bo boholo kapa bo bonyenyane, kapa ka linako tse ling lihlopha tse ikhethileng tse matlafatsang le lihlopha tse sa ikhethileng tsa matlafatso lia etsahala.

Tlhahiso ea A-1

■ Lintlha tse qalang tsa semmuso

——Re-design ea khale.

■ Khakanyo ea semmuso e phahame haholo —— Ho eketsa mocheso oa denaturation hantle le ho lelefatsa nako ea denaturation.

Mofuta oa A-2 Mg²⁺ ho tsepamisa maikutlo

■ Mokha oa Mg²⁺ mahloriso a phahame haholo -— Fokotsa hantle mohopolo oa Mg2 +: Lokisetsa Mg²⁺ mahloriso ka letoto la likarabelo ho tloha ho 1 mM ho isa ho 3 mM ka karohano ea 0.5 mM ho fumana Mg²⁺ mahloriso bakeng sa template le primer ka 'ngoe.

A-3 Thermostable polymerase

■ Palo ea enzyme e fetelletseng —— Fokotsa palo ea enzyme ka nepo ka linako tse ling tsa 0.5 U.

Mocheso oa ho koala oa A-4

■ Mocheso o kenyang letsoho o tlase haholo —— Ho eketsa mocheso oa ho koala ka nepo kapa ho sebelisa mokhoa oa mekhahlelo e 'meli

Lipotoloho tsa A-5 PCR

■ Lipotoloho tse ngata tsa PCR ——Ho fokotsa palo ea lipotoloho tsa PCR.

P: Lihlopha tse hlabang kapa tse tlotsang

Tlhahiso ea A-1—— Boemo bo ikgethang ba maemo a mabe— - Iketsetse meralo ea mantlha, fetola boemo le bolelele ba semmuso ho ntlafatsa botumo ba eona; kapa ho etsa sehlaha sa PCR.

DNA ea A-2

——Thempleite ha e ea hloeka— - Hloekisa template kapa u ntše DNA e nang le lisebelisoa tsa tlhoekiso.

EA-3 Mg²⁺ ho tsepamisa maikutlo

——Mahl²⁺ ho tsepamisa mohopolo ho phahame haholo -— Fokotsa hantle Mg²⁺ mahloriso: Lokisetsa Mg²⁺ mahloriso ka letoto la likarabelo ho tloha ho 1 mM ho isa ho 3 mM ka karohano ea 0.5 mM ho fumana Mg²⁺ mahloriso bakeng sa template le primer ka 'ngoe.

E-4 dNTP

——Khokahano ea li-dNTP li holimo haholo —— Fokotsa mohopolo oa dNTP ka nepo

Mocheso oa Annealing oa 5

——Thempereichara e tlaase e kenang ——Ho eketsa hantle mocheso o tlatsetsang

Lipotoloho tsa A-6

—Kenale palo ea potoloho

P: Ke template ea bokae ea DNA e lokelang ho kenyelletsoa ho 50 μl PCR reaction system?

P: U ka holisa likaroloana tse telele joang?

Mohato oa pele ke ho khetha polymerase e loketseng. Kamehla Taq polymerase e ke ke ea bala liphoso ka lebaka la ho haelloa ke ts'ebetso ea 3'-5 'ea tokollo,' me ho se tšoane hantle ho tla fokotsa ts'ebetso ea likhechana. Ka hona, Taq polymerase e tloaelehileng e ke ke ea holisa hantle likaroloana tsa sepheo tse kholo ho feta 5 kb. Taq polymerase e nang le phetoho e khethehileng kapa botšepehi bo phahameng ba polymerase e lokela ho khethoa ho ntlafatsa katleho le ho fihlela litlhoko tsa sehlotšoana se selelele. Ntle le moo, kholiso ea likhechana tse telele e boetse e hloka phetoho e tsamaellanang ea moralo oa khale, nako ea denaturation, nako ea katoloso, pH ea buffer, jj. Hangata, li-primers tse nang le 18-24 bp li ka lebisa ho chai e ngata. Bakeng sa ho thibela tšenyo ea template, nako ea denaturation ho 94 ° C e lokela ho fokotsoa ho isa ho 30 sec kapa ka tlase ho potoloho, mme nako ea ho nyoloha mocheso ho ea ho 94 ° C pele amplification e lokela ho ba tlase ho 1 min. Ho feta moo, ho beha mocheso oa katoloso ka 68 ° C le ho rala nako ea katoloso ho latela sekhahla sa 1 kb / min ho ka netefatsa kholiso e sebetsang ea likhechana tse telele.

Q: U ka ntlafatsa ts'epahalo ea kholiso ea PCR joang?

Sekhahla sa phoso ea kholiso ea PCR se ka fokotsoa ka ho sebelisa li-polymerase tse fapaneng tsa DNA ka bots'epehi bo phahameng. Har'a li-polymerase tsohle tsa Taq DNA tse fumanehang ho fihlela joale, enzyme ea Pfu e na le sekhahla se tlase sa phoso le botšepehi bo phahameng ho fetisisa (bona tafole e hoketsoeng). Ntle le khetho ea enzyme, bafuputsi ba ka tsoela pele ho fokotsa sekhahla sa phetoho ea PCR ka ho ntlafatsa maemo a karabelo, ho kenyeletsoa le ho ntlafatsa sebopeho sa buffer, polasetiki ea thermostable polymerase le ho ntlafatsa palo ea potoloho ea PCR.

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