I-Hot-start Taq enzyme isetshenziswa kabanzi.Uma kuqhathaniswa ne-DNA polymerase evamile, i-enzyme ye-Taq eqala ukushisa ingagwema ngempumelelo okunye ukukhulisa okungaqondile kanye nokwakheka kwama-primer dimers, futhi ingathuthukisa ngempumelelo izinga lokuphumelela lokukhulisa isakhi sofuzo.Ikakhulukazi emkhakheni wokuhlolwa kofuzo, i-enzyme ye-Taq eshisayo ikhonjwe njengendinganiso eyisibopho embonini, futhi i-DNA polymerase evamile akufanele isetshenziswe.Njengoba kubonakala kulokhu okungenhla, ama-enzyme e-Taq aqala ashisayo asetshenziswa kakhulu.Njengamanje, kunezinhlobo eziningi zama-enzyme e-Taq aqala ashisayo emakethe yasekhaya, kodwa awekho ama-enzyme e-Taq ashisayo amaningi anekhwalithi ephezulu.Sibhekene nemikhiqizo eminingi ye-enzyme ye-Taq eshisayo, kufanele sikhethe kanjani?

1. Khetha i-enzayimu ye-Taq enesiqalo esishisayo enekhono eliphezulu lokukhulisa

Ukusebenza kahle kokukhulisa i-PCR kuhlobene eduze nokusebenza kwe-Taq enzyme.Ngemuva kokuthi uhlelo oluhle lokusabela lwe-enzyme ye-Taq seluthuthukisiwe, ukusebenza kahle kokukhulisa kungaphezu kwama-95%, futhi ububanzi bokukhulisa inani lethempulethi yokuqala bubanzi.Ukukhulisa okwanelisayo kungatholwa uma okuqukethwe kofuzo okuqondiwe kuphansi, futhi akulula ukungeniswa ushevu lapho inani lesifanekiso liphezulu, futhi isikhathi sokukhulisa i-exponential side.Ku-enzyme ye-Taq engasebenzi kahle, ngisho noma isistimu yokusabela ithuthukiswe izikhathi eziningi, ukusebenza kahle kokukhulisa kusengaphansi kwama-90%, ukwakheka kwe-"S" kwejika lokukhulisa akubonakali, umthambeka mncane, futhi ijika liyisicaba.Uma inani lesifanekiso liphansi, alikwazi ukukhuliswa, futhi uma inani lesifanekiso liphezulu, umphumela wokukhulisa awulungile.Ngakho-ke, ukukhethwa kwama-polymerase e-DNA anekhono eliphezulu lokukhulisa i-amplification kubalulekile empumelelweni ye-PCR ne-qPCR.

2. Khetha i-enzyme ye-Taq eshisayo enamandla anamandla e-enzyme

Amandla enzymatic we-Taq enzyme ahlobene nokusebenza kahle kokukhulisa.Ngokuvamile, lapho amandla enzymatic eqala ukushisa i-enzyme ye-Taq eqinile, isikhathi sokukhula komchazi we-PCR siba side, ijika elijwayelekile 'elimise okwe-S', liyakhuphuka inani lesignali ye-fluorescence, futhi kufaneleka kakhulu ukutholwa kwe-multiplex PCR.Ama-polymerase e-Brand DNA anamandla enzymatic abuthakathaka ngokuvamile angasekela kuphela ukusabela okungu-2-plex.Uma wenza ukusabela okungu-3-plex, ijika lokukhulisa liphansi, inani lesignali ye-fluorescence liphansi, futhi alikho ijika elivamile lokukhulisa, ngakho-ke imiphumela kunzima ukuyihlulela.

3. Khetha i-enzayimu ye-Taq enesiqalo esishisayo ezwela kakhulu

Ngokuvamile, i-DNA polymerase inekhono eliphezulu lokukhulisa i-amplification kanye nokuzwela okuphezulu, kodwa kukhona nokungahambisani.Uma ubuningi bofuzo oluqondiwe lwesampula okufanele lukhuliswe buphansi, kuyanconywa ukuhlola ukuzwela kokukhulisa i-Taq enzyme.Indlela ejwayeleke kakhulu yokuthola iwukwenza ukuhlanjululwa kwe-gradient okuphindwe ka-10 noma okuphindwe ka-5 kocezu lofuzo oluqondiwe lwe-plasmid, ukwenza ukutholwa kwe-PCR ekuhlanjululweni okuphansi, bese ukhetha i-enzyme ye-Taq eqala ukushisa enokuzwela okuphezulu kokutholwa.

Kungabonakala kulokhu okungenhla ukuthi abacwaningi badinga ukukhetha ngokuvumelana nezidingo zabo zokuhlola kanye nezimo zoxhaso.Kungcono ukwenza isilingo sokukhulisa i-gradient dilution ukuze kutholwe ukusebenza kahle kokukhulisa nokuzwela kwe-enzyme ye-Taq eqala ukushisa.

Isibonelo seForegene'I-Taq DNA Polymerase:

I-Foreasy HS Taq DNA Polymerase

Incazelo

I-Foreasy HS Taq DNA Polymerase iyi-DNA polymerase evezwa kubhaktheriya yobunjiniyela be-Escherichia coli ngobuchwepheshe bokuhlanganiswa kabusha kwezakhi zofuzo.I-enzayimu ihlanganiswe ne-buffffer yokusabela eyingqayizivele, eyenza umkhiqizo umelane kakhulu futhi uhambisane, futhi ingasebenzisa ngokuqondile isampula ye-lysate (isistimu ye-Forgene Lysis) njengesifanekiso sokusabela kokutholwa.

Isicelo

I-PCR efanelekile kanye nokutholwa kwe-PCR yobuningi yezifanekiso ezicwengiwe nezifanekiso ezingacwengisisiwe.

Ikhwalithi yokulawula

1.Awukho umsebenzi we-nuclease exogenous otholiwe

2.Indlela ye-PCR yokuthola i-DNA ye-genomic eyinsalela ngaphandle komsingathi

3.Ingakwazi ukukhulisa ngempumelelo ufuzo lwekhophi eyodwa kuGenome yomuntu

4.Gcina ekamelweni lokushisa isonto lonke,umsebenzi ongabonakali uyashintsha

Imininingwane yomkhiqizo: https://www.foreivd.com/foreasy-hs-taq-dna-polymerase-product/