Hot-start Taq enzyme imagwiritsidwa ntchito kwambiri.Poyerekeza ndi DNA polymerase wamba, enzyme ya Taq yotentha imatha kupewa kukulitsa kwina kosadziwika bwino komanso kupanga ma primer dimers, ndipo imatha kusintha bwino kuchuluka kwa jini.Makamaka pankhani yoyezetsa majini, enzyme ya Taq yotentha yadziwika kuti ndiyofunikira pamakampani, ndipo DNA polymerase wamba sayenera kugwiritsidwa ntchito.Monga tawonera pamwambapa, ma enzymes a Taq otentha amagwiritsidwa ntchito kwambiri.Pakadali pano, pali mitundu yambiri ya ma enzymes a Taq oyambira pamsika wapanyumba, koma palibe ma enzymes ambiri a Taq omwe ali ndipamwamba kwambiri.Poyang'anizana ndi zinthu zambiri zoyambira za Taq enzyme, tiyenera kusankha bwanji?

1. Sankhani puloteni ya Taq yotentha yokhala ndi mphamvu zokulitsa

Kuchita bwino kwa PCR kumayenderana kwambiri ndi magwiridwe antchito a Taq enzyme.Pambuyo pokonzekera bwino Taq enzyme reaction system, mphamvu yokulitsa imakhala yopitilira 95%, ndipo kuchuluka kwa kuchuluka kwa template yoyambira ndikokulirapo.Kukulitsa kokhutiritsa kungapezeke pamene chandamale cha jini chili chochepa, ndipo sikophweka kukhala ndi poizoni pamene chiwerengero cha template chili chochuluka, ndipo nthawi yokulitsa chidziwitso ndi yaitali.Kwa enzyme ya Taq yokhala ndi vuto losagwira bwino ntchito, ngakhale machitidwe amachitidwe asinthidwa nthawi zambiri, mphamvu yokulitsa imakhalabe yochepera 90%, mawonekedwe a "S" amplification curve sizodziwikiratu, otsetsereka ndi ochepa, ndipo phirilo ndi lathyathyathya.Pamene chiwerengero cha template chili chochepa, sichikhoza kukulitsidwa, ndipo pamene kuchuluka kwa template kuli kwakukulu, zotsatira zokulitsa sizili bwino.Chifukwa chake, kusankha ma DNA ma polymerases okhala ndi matalikidwe apamwamba ndikofunikira kuti PCR ndi qPCR zichite bwino.

2. Sankhani enzyme ya Taq yotentha yokhala ndi mphamvu yamphamvu ya enzyme

Mphamvu ya enzymatic ya Taq enzyme imakhudzana ndi kukulitsa bwino.Nthawi zambiri, mphamvu ya enzymatic ya enzyme ya Taq yotentha kwambiri, ndi nthawi yotalikirapo ya kukula kwa PCR, kupindika kwa 'S-mawonekedwe a S', kuchuluka kwa siginecha ya fluorescence, komanso koyenera kuzindikira ma PCR ambiri.Ma polymerases a Brand DNA okhala ndi mphamvu yofooka ya enzymatic amatha kuthandizira machitidwe a 2-plex.Mukamachita ma 3-plex reaction, curve yokulitsa imakhala yotsika, mtengo wa siginecha ya fluorescence ndi wotsika, ndipo palibe mayendedwe okulirapo, kotero zotsatira zake zimakhala zovuta kuweruza.

3. Sankhani enzyme ya Taq yotentha yokhala ndi chidwi chachikulu

Nthawi zambiri, DNA polymerase imakhala ndi mphamvu zambiri zokulitsa komanso kumva bwino, koma palinso zosagwirizana.Ngati kuchuluka kwa jini yachitsanzo kuti ikulitsidwe kuli kochepa, tikulimbikitsidwa kuyesa kukhudzika kwa Taq enzyme.Njira yodziwika kwambiri yodziwira ndikutulutsa kachulukidwe ka 10 kapena ka 5 kagawo kakang'ono ka jini ya plasmid, kuzindikiritsa PCR kumunsi kwa dilution, ndikusankha enzyme ya Taq yotentha yokhala ndi chidwi chodziwikiratu.

Zitha kuwoneka kuchokera pamwambazi kuti ochita kafukufuku akuyenera kusankha malinga ndi zomwe akufuna komanso momwe angathandizire ndalama.Ndibwino kuti muyesere kuyesa kwa gradient dilution kuti muwone kukwera bwino komanso kukhudzika kwa enzyme ya Taq yotentha.

Chitsanzo cha Foregene's Taq DNA Polymerase:

Foreasy HS Taq DNA Polymerase

Kufotokozera

Foreasy HS Taq DNA Polymerase ndi DNA polymerase yofotokozedwa mu Escherichia coli engineering bacteria pogwiritsa ntchito gene recombination technology.Enzyme imaphatikizidwa ndi buffffer yapadera, yomwe imapangitsa kuti mankhwalawa akhale osagwirizana komanso ogwirizana, ndipo amatha kugwiritsa ntchito mwachindunji chitsanzo cha lysate (Foregene Lysis system) ngati template kuti azindikire.

Kugwiritsa ntchito

Ubwino wa PCR ndi kuchuluka kwa PCR kuzindikira kwa ma tempulo oyeretsedwa ndi ma tempulo osayeretsedwa.

Kuwongolera Kwabwino

1.Palibe zochitika zamtundu wa nuclease zomwe zapezeka

2.PCR njira yodziwira DNA yotsalira ya genomic popanda wolandira

3.Itha kukulitsa bwino jini yamtundu umodzi mu Genome yamunthu

4.Sungani kutentha kwa firiji kwa sabata, zochitika zosaoneka zimasintha

Zambiri zamalonda: https://www.foreivd.com/foreasy-hs-taq-dna-polymerase-product/