QuickEasyᵀᴹ Nako ea 'Nete PCR Kit-Taqman
Tlhaloso ea Kit:
◮Sistimi e ikhethileng ea PCR e etsa 2×QuickEasyTMReal PCR Mix-Taqman e tsamaellana haholoanyane.
◮Hot-start Foregene HS Taq Polymerase e na le bokhoni bo phahameng ba ho holisa, kutlo e phahameng ea amplification, le ho hlaka ho hoholo ho holimo.
◮2 × QuickEasyTMReal PCR Mix -Taqman e sebelisa sistimi e ikhethileng e ntlafalitsoeng ke Foregene ho ntlafatsa kutlo le ho hlaka ha tlhahlobo e ikhethileng ea tatellano, e ka sebelisoang ho etsa qeto ea ho fapana ha linomoro tsa genotyping le ho kopitsa le ho fumana liphetho tse nepahetseng.
◮Sehlahisoa sena se tla le dae ea ka hare ea ROX, e ka sebelisoang ho tlosa semelo sa matšoao le phoso ea matšoao lipakeng tsa liliba, tse loketseng bareki ho li sebelisa ka mefuta e fapaneng ea lisebelisoa tsa PCR.
Litlhaloso
2 × QuickEasyTMReal PCR Mix-Taqman e fanoe ke QuickEasyTMReal Time PCR Kit-Taqman kit ke sistimi e ncha e kopanyang e ncha e etselitsoeng karabelo ea amplification ea Real Time PCR e sebelisa li-fluorescent probes.Setsi sa eona sa mantlha sa Foregene HS Taq DNA Polymerase se ipapisitse le phetoho ea Antibody, e sebelisoang hammoho le sistimi ea Foregene e ntlafalitsoeng ka ho fetesisa ea PCR, e na le ts'ebetso e phahameng haholo le matlafatso.Ha e bapisoa le motsoako o tloaelehileng oa PCR, e na le bokhoni bo matla ba ho holisa le sekhahla se tlase sa ho se lumellane.E ka sebelisoa ho fluorescence quantitative PCR reaction ho fokotsa amplification e seng e khethehileng le ho ntlafatsa ho nepahala ha PCR.E ka ntlafatsa haholo boleng ba sehlahisoa le kutloisiso ea karabelo.Ka nako e ts'oanang, ROX e ile ea fanoa e le dae ea ka hare ea litšupiso.
The QuickEasyTMNako ea 'nete ea PCR Kit-Taqman kit e na le sebaka se pharaletseng sa ts'ebeliso ho palo ea template,' me e ka fumana sekhahla se setle libakeng tse fapaneng tsa palo, 'me e khona ho lekanya le ho bona mefuta ea liphatsa tsa lefutso ka ho pheta-pheta hantle le kholiseho e phahameng.
Litlhaloso
Litokisetso tse 200,Litokisetso tse 500,Litokisetso tse 1000,2000 Preps
Likarolo tsa Kit
| QuickEasyTMNako ea 'Nete PCR Kit-Taqman(20μltsamaiso) | ||||
| Likahare tsa Kit | QP-01121 | QP-01122 | QP-01123 | QP-01124 |
| 200 Preps | 500 Preps | 1000 Preps | 2000 Preps | |
| 2×Ka potlakoBonoloTM PCR ea 'nete Kopanya- Taqman | 1 mL×2 | 1.7 mL×3 | 1.7 mL×6 | 1.7 mL×12 |
| 20×ROX Reference Dae | 200μL | 0.5 mL | 1 mL | 1 mL×2 |
| DNase-Free ddH2O | 1.7 mL | 1.7 mL×2 | 10 ml | 20 ml |
| IFU | 1sengoathoana | 1sengoathoana | 1sengoathoana | 1sengoathoana |
Likarolo le melemo
■ Sistimi e ikhethang ea ntlafatso ea PCR e etsa 2×QuickEasyTMReal PCR Mix-Taqman e tsamaellana haholoanyane.
■ Hot-start Foregene HS Taq Polymerase e na le matla a holimo a ho holisa, a snetivity e phahameng ea amplification, le ho hlaka ho holimo ho matla.
■ 2× QuickEasyTM Real PCR Mix -Taqman e sebelisa sistimi e ikhethileng e ntlafalitsoeng ke Foregene ho ntlafatsa kutlo le ho hlaka ha tlhahlobo e ikhethileng ea tatellano, e ka sebelisoang ho etsa qeto ea ho fapana ha linomoro tsa genotyping le ho kopitsa le ho fumana liphetho tse nepahetseng.
■ Sehlahisoa sena se tla le dae ea ka hare ea ROX, e ka sebelisoang ho felisa semelo sa matšoao le phoso ea pontšo pakeng tsa liliba, e leng bonolo hore bareki ba e sebelise ka mefuta e fapaneng ea lisebelisoa tsa PCR tse ngata.
Sesebelisoa sa Kit
■ Fluorescence quantitative PCR bakeng sa tlhahlobo ea palo ea template;
■ Cntlafatso ea PCR e sa tloaelehang;
■ E ka sebelisoa ho lemoha allele.
Storage le Shelf bophelo
Sekoti se lokela ho bolokoa ho -20 ° C lefifing;haeba e sebelisoa khafetsa, e ka boela ea bolokoa ho 4 ° C ka nako e khutšoanyane (sebelisa pele ho matsatsi a 10).
Melao-motheo ea moralo oa nako ea 'nete ea PCR
Pele ho Primer le Reverse Primer
Bakeng sa nako ea 'nete ea PCR, moralo oa primer o bohlokoa haholo.Li-primers li amana le ho hlaka le katleho ea ntlafatso ea PCR, 'me li ka etsoa ho ipapisitsoe le melaoana e latelang:
Bolelele ba pele: 18-30bp.
Likahare tsa GC: 40-60%.
Tm boleng: Software ea moralo oa Primer, joalo ka Primer 5, e ka fana ka boleng ba Tm ba primer.Litekanyetso tsa Tm tsa li-primers tse holimo le tse tlase li lokela ho ba haufi ka hohle kamoo ho ka khonehang.Mokhoa oa ho bala oa Tm o ka boela oa sebelisoa: Tm = 4 °C (G + C) + 2 °C (A + T).Ha o etsa PCR, mocheso o ka tlase ho boleng ba pele ba Tm oa 5 ° C hangata o khethoa joalo ka mocheso o kenang (keketseho e lumellanang ea mocheso oa annealing e ka eketsa boits'oaro ba PCR).
Li-Primers le lihlahisoa tsa PCR:
Bolelele ba sehlahisoa sa ntlafatso ea PCR ka ho khetheha ke 100-150bp.
Li-primer tsa moralo sebakeng sa bobeli sa sebopeho sa template li lokela ho qojoa ka hohle kamoo ho ka khonehang.
Qoba ho theha metheo e 2 kapa ho feta e tlatselletsanang lipakeng tsa 3′ ea lithako tse holimo le tse tlase.
Primer 3′ terminal base e ka se be teng ka tse ling tse 3 tse latellanang tsa G kapa C.
Li-primers ka botsona li ke ke tsa ba le meaho e tlatselletsang, ho seng joalo sebopeho sa hairpin se tla thehoa, se amang ntlafatso ea PCR.
ATCG e lokela ho ajoa ka ho lekana ka hohle kamoo ho ka khonehang ka tatellano ea pele, 'me setsi sa 3′ se lokela ho qojoa joalo ka T.
Sehlomathiso sa 1: Sephutheloana sa tlatsetso sa Cell Direct RT-qPCR Kit
1.Seleng Lysis Tharollo
|
| |||
| Likarolo tsa Kit (Sistimi ea lysis ea seliba tse 24 / seliba) | DRT-01011-A1 | DRT-01011-A2 | |
| 100 T | 500 T | ||
| KaroloI | Buffer CL | 20 ml | 100 ml |
| Foregene Protease Plus II | 400 μl | 1 ml × 2 | |
| Buffer ST | 1 ml × 2 | 10 ml | |
| KaroloII | DNA Raba | 400 μl | 1 ml × 2 |
2.RT Motsoako
|
| |
| Likarolo tsa Kit (Sistimi ea karabelo ea 20 μl) | DRT-01011-B1 |
| 200 T | |
| 5 × Direct RT Mix | 800 μl |
| RNase-Free ddH2O | 1.7 ml × 2 |
|
| ||
| Likarolo tsa Kit (Sistimi ea karabelo ea 20 μl) | DRT-01011-C1 | DRT-01011-C2 |
| 200 T | 1000 T | |
| 2 × Direct qPCR Mix-SYBR | 1 ml × 2 | 1.7 ml × 6 |
| 50 × ROX Reference Dye | 40 μl | 200 μl |
| RNase-Free ddH2O | 1.7 ml | 10 ml |
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