Bst 2.0 DNA Polymerase(Glycerol free)
Ko te Bst DNA polymerase V2 i ahu mai i te Bacillus stearothermophilus DNA Polymerase I, he 5′→3′ te mahi DNA polymerase me te mahi whakakapi mekameka kaha, engari kaore he mahi 5′→3′ exonuclease.Ko te Bst DNA Polymerase V2 he mea pai mo te whenu-displacement, te whakakaha hauwaro i te RAMA (Loop mediated isothermal amplification) me te raupapa tere.
Waehanga
| Waehanga | HC5005A-01 | HC5005A-02 | HC5005A-03 |
| BstDNApolymerase V2(Glycerol-free)(8U/μL) | 0.2 mL | 1 mL | 10 mL |
| 10×HC Bst V2 Kaipupuri | 1.5 mL | 2×1.5 mL | 3×10 mL |
| MgSO4(100mM) | 1.5 mL | 2×1.5 mL | 2×10 mL |
Nga tono
1.LAMP whakakaha hauwaro
2.DNA whenu tauhohenga nekehanga kotahi
3.High GC raupapa ira
4.DNA raupapa o te taumata nanogram.
Tikanga Rokiroki
Te kawe i raro i te 0°C ka penapena ki te -25°C~-15°C.
Whakamaramatanga Waeine
Kotahi te waeine kua tautuhia ko te nui o te whākōkī e whakauru ana i te 25 nmol o te dNTP ki roto i te waikawa waikawa i roto i te 30 meneti i te 65°C.
Mana Kounga
1.Te Whakatairanga Maamaa Protein (SDS-PAGE):Ko te ma o Bst DNA polymerase V2 ko ≥99% kua whakatauhia e te tātaritanga SDS-PAGE ma te whakamahi i te kitenga Coomassie Blue.
2.Mahi Exonuclease:Ko te werohanga o te tauhohenga 50 μL kei roto te iti rawa o te 8 U o te Bst DNA polymerase V2 me te 1 μg λ -Hind Ⅲ digest DNA mo nga haora 16 i te 37 ℃ ka kore e kitea te paheketanga ka kitea.
3.Mahi Nickase:Ko te werohanga o te tauhohenga 50 μL kei roto te iti rawa o te 8 U o te Bst DNA polymerase V2 me te 1 μg pBR322 DNA mo nga haora 16 i te 37°C ka kore e kitea te paheketanga kua whakatauhia.
4.Mahi RNase:Ko te werohanga o te tauhohenga 50 μL kei roto te iti rawa o te 8 U o te Bst DNA polymerase V2 me te 1.6 μg MS2 RNA mo nga haora 16 i te 37°C ka kore e kitea te paheketanga kua whakatauhia.
5.E. coli DNA:Ko te 120 U o te Bst DNA polymerase V2 ka tirotirohia mo te aroaro o te E. coli genomic DNA ma te whakamahi i te TaqMan qPCR me nga tohu tuatahi mo te waahi E. coli 16S rRNA.Ko te pokenga DNA genomic E. coli he ≤1 Tārua.
Tauhohenga RAMA
| Waehanga | 25μL |
| 10×HC Bst V2 Kaipupuri | 2.5 μL |
| MgSO4 (100mM) | 1.5 μL |
| dNTP (10mM ia) | 3.5 μL |
| SYTO™ 16 Kākāriki (25×)a | 1.0 μL |
| Te ranunga tuatahib | 6 μL |
| Bst DNA Polymerase V2 (Glycerol-free) (8 U/uL) | 1 μL |
| tauira | × μL |
| ddH₂O | Tae atu ki te 25 μL |
Tuhipoka:
1) a.SYTOTM 16 Kākāriki (25×): E ai ki nga hiahia whakamatautau, ka taea te whakamahi etahi atu waikano hei whakakapi;
2) b.Ranu tuatahi: ka whiwhi ma te ranu 20 µ M FIP, 20 µ M BIP, 2.5 µ M F3, 2.5 µ M B3, 5 µ M LF, 5 µ M LB me etahi atu pukapuka.
Tauhohenga me te Tikanga
1 × HC Bst V2 Buffer, kei waenganui i te 60°C me te 65°C te pāmahana heki.
Te Wera Whakakore
80 °C, 20 meneti
