Foreasy Taq DNA Polymerase
Kit Description:
◮Yakanyanya kujeka: Iyo enzyme ine imwe inopisa-yekutanga chiitiko.
◮Kukurumidza Kuwedzera: 10 sec/kb.
◮Yakanyanya kuchinjika template: inogona kushandiswa kunyatso kusimudzira GC Yakakwirira kukosha, akasiyana-siyana akaoma-ku-kurisa DNA template.
◮Kuvimbika kwakasimba: yakajairwa Taq Enzyme 6 nguva.
◮Kugadzikana kwekushisa kwakasimba: Inogona kuiswa pa 37 ° C kwevhiki uye inochengetedza zvinopfuura 90% chiitiko.
Tsanangudzo
Foreasy Taq DNA Polymerase itsva Taq enzyme inoratidzwa muEscherichia coli engineering bhakitiriya nejene recombination tekinoroji.Iyo enzyme pachayo ine imwe inopisa-yekutanga basa uye inogona kushandiswa kune yakajairika PCR uye qPCR;ine 5'→ 3' DNA polymerase chiitiko uye 5'→ 3' exonuclease chiitiko, asi hapana 3'→ 5' exonuclease chiitiko.
Kit zvikamu
| Chikamu | IM-01011 | IM-01012 | IM-01013 |
| Foreasy Taq DNA Polymerase(5 U/μL) | 5000 U (1 mL) | 50 KU (10 mL) | 500 KU (100 mL) |
| 2 × Taq Reaction Buffer | 25 mL ×5 | 250 mL ×5 | 500 mL × 25 |
Zvimiro&zvakanakira
- Yakanyanya kujeka: Iyo enzyme ine imwe inopisa-yekutanga chiitiko.
-Kukurumidza Kuwedzera: 10 sec/kb.
-Yakanyanya kuchinjika template: inogona kushandiswa kunyatso kusimudzira GC Yakakwirira kukosha, akasiyana-siyana akaoma-ku-kurisa DNA template.
- Kuvimbika kwakasimba: yakajairwa Taq Enzyme katanhatu.
- Yakasimba yekudziya kugadzikana: Inogona kuiswa pa37 ° C kwevhiki uye inochengetedza zvinopfuura 90% chiitiko.
Kit application
Yakasiyana PCR/qPCR masisitimu uye yakananga PCR masisitimu
PCR kukwidziridzwa kweDNA zvimedu
DNA label
DNA kutevedzana
PCR A-muswe
U Definition
1U: Huwandu hwe enzyme inodiwa kuisa 10 nmol ye deoxynucleotides mu acid-insoluble matter uchishandisa activated salmon sperm DNA se template / primer kwemaminitsi makumi matatu pa 74 ° C.
Reaction Condition
| Tembiricha | Nguva | Cycle |
| 37°C | 5 mins | 1 |
| 94°C | 5 mins | 1 |
| 94°C | 10 Secs | 35 |
| 60°C | 10 Secs | |
| 72°C | 20 sec/kb | |
| 72°C | 2mins | 1 |
Storage
-20 ± 5 °C kwemakore maviri kana -80 °C kwekuchengetedza kwenguva refu.
Hapana masaini ekuwedzera
1.Taq DNA Polymerase mukiti inorasikirwa nebasa rayo nekuda kwekuchengetedza kusina kunaka kana kupera kwekiti.
Kurudziro: Simbisa mamiriro ekuchengetedza ekiti;wedzerazve huwandu hwakakodzera hweTaq DNA Polymerase kuPCR system kana kutenga itsva Real Time PCR Kit yezviedzo zvinoenderana.
2.Kune akawanda inhibitors yeTaq DNA Polymerase muDNA template.
Zvaungaita: Natsazve template kana kuderedza huwandu hwetemplate yakashandiswa.
3.Mg2 + concentration haina kukodzera.
Kurudziro: Iyo Mg2 + yekumisikidza ye2 × Real PCR Mix yatinopa ndeye 3.5mM.Nekudaro, kune mamwe akakosha ekutanga uye matemplate, iyo Mg2 + yekumisikidza inogona kunge yakakwira.Naizvozvo, unogona kuwedzera zvakananga MgCl2 kukwidziridza iyo Mg2+ concentration.Zvinokurudzirwa kuwedzera iyo Mg2+ 0.5mM nguva yega yega kuitira optimization.
4.Iyo PCR amplification mamiriro haana kukodzera, uye kutevedzana kweprimer kana kuisa pfungwa hakuna kunaka.
Mazano: simbisa kururamisa kwekutevedzana kweprimer uye primer haina kuderedzwa;kana chiratidzo chekusimudzira chisina kunaka, edza kudzikisira tembiricha yekumisa uye gadzirisa iyo primer concentration nenzira kwayo.
5.Kuwanda kwe template kuduku kana kuwanda.
Kurudziro: Ita template linearization gradient dilution, uye sarudza iyo template yekumisikidza ine yakanakisa PCR mhedzisiro yeChaiyo Nguva PCR kuyedza.
NTC ine yakawandisa fluorescence kukosha
1.Reagent kusvibiswa kunokonzerwa panguva yekushanda.
Kurudziro: Tsiva nema reagents eChaiyo Nguva PCR kuyedza.
2.Kusvibiswa kwakaitika panguva yekugadzirira kwePCR reaction system.
Kurudziro: Tora matanho ekudzivirira anodiwa panguva yekushanda, senge: kupfeka magurovhosi e latex, kushandisa piipi tipi ine sefa, nezvimwe.
3.The primers yakasvibiswa, uye kuderedzwa kwezvinyorwa kuchaita kuti kusava kwakananga amplification.
Kurudziro: Shandisa SDS-PAGE electrophoresis kuona kana maprimers akashatiswa, woatsiva nemaprimer matsva eChaiyo Nguva PCR kuyedza.
Primer dimer kana kuti non-specific amplification
1.Mg2 + concentration haina kukodzera.
Kurudziro: Iyo Mg2 + yepakati ye2 × Real PCR EasyTM Mix yatinopa ndeye 3.5 mM.Nekudaro, kune mamwe akakosha ekutanga uye matemplate, iyo Mg2 + yekumisikidza inogona kunge yakakwira.Naizvozvo, unogona kuwedzera zvakananga MgCl2 kukwidziridza iyo Mg2+ concentration.Zvinokurudzirwa kuwedzera iyo Mg2+ 0.5mM nguva yega yega kuitira optimization.
2.The PCR annealing tembiricha yakaderera.
Zano: Wedzera tembiricha yePCR annealing ne1 ℃ kana 2 ℃ nguva yega yega.
3.Chigadzirwa chePCR chakareba.
Kurudziro: Hurefu hweiyo Real Time PCR chigadzirwa hunofanirwa kunge huri pakati pe100-150bp, kwete kupfuura 500bp.
4.The primers yakasvibiswa, uye kusvibiswa kwezvinyorwa zvinotungamirira pakuonekwa kwekuwedzerwa kwakananga.
Kurudziro: Shandisa SDS-PAGE electrophoresis kuona kana maprimers akashatiswa, woatsiva nemaprimer matsva eChaiyo Nguva PCR kuyedza.
5.Iyo PCR system haina kunaka, kana hurongwa hwakanyanya kuduku.
Zano: Iyo PCR reaction system idiki zvakanyanya inokonzeresa kuti yekuona iderere.Zvakanakisa kushandisa reaction system inokurudzirwa neakawanda PCR chiridzwa kumhanyisa zvakare Yechokwadi Nguva PCR kuyedza.
Kusagona kudzokororwa kwehuwandu hunokosha
1.Chiridzwa chiri kutadza kushanda.
Zano: Panogona kunge paine kukanganisa pakati pegomba rega rega rePCR yechiridzwa, zvichikonzera kusaberekana zvakanaka panguva yekutarisira tembiricha kana kuona.Ndokumbira utarise maererano nemirairo yechiridzwa chinoenderana.
2.Kuchena kwemuenzaniso hakuna kunaka.
Kurudziro: Samples dzisina kuchena dzinotungamira mukutadza kuberekana kwekuyedza, izvo zvinosanganisira kuchena kwetemplate uye yekutanga.Zvakanakisa kuchenesa template, uye iyo yekutanga inonatswa zvakanyanya neSDS-PAGE.
3.Iyo PCR system yekugadzirira uye nguva yekuchengetedza yakanyanyisa.
Kurudziro: Shandisa iyo Real Time PCR system yePCR kuyedza nekukurumidza mushure mekugadzirira, uye usaisiye parutivi kwenguva yakareba.
4.Iyo PCR amplification mamiriro haana kukodzera, uye kutevedzana kweprimer kana kuisa pfungwa hakuna kunaka.
Mazano: simbisa kururamisa kwekutevedzana kweprimer uye primer haina kuderedzwa;kana chiratidzo chekusimudzira chisina kunaka, edza kudzikisira tembiricha yekumisa uye gadzirisa iyo primer concentration nenzira kwayo.
5.Iyo PCR system haina kunaka, kana hurongwa hwakanyanya kuduku.
Zano: Iyo PCR reaction system idiki zvakanyanya inokonzeresa kuti yekuona iderere.Zvakanakisa kushandisa reaction system inokurudzirwa neakawanda PCR chiridzwa kumhanyisa zvakare Yechokwadi Nguva PCR kuyedza.
