Bst 2.0 DNA Polymerase (Glycerolum gratis)
Bst DNA polymerasis V2 derivatur ab Bacillo stearothermophilo DNA Polymerase I, quod actio polymerasis 5′ →3′ et catena valida postea activitatem exonucleasem habet, sed non est 5′ →3′ exonucleasis actio.Bst DNA Polymerase V2 optime convenit ad locum subductionis, amplificationis isothermal LAMPIS (Loop amplificationis isothermal mediatae) et celeris sequelae.
Components
| Component | HC5005A-01 | HC5005A-02 | HC5005A-03 |
| BstDNApolymerase V2(Glycerolum-free)(8U/μL) | 0.2 mL | 1 mL | 10 mL |
| 10×HC Bst V2 Buffer | 1.5 mL | 2×1.5 mL | 3×10 mL |
| MgSO4(100mM) | 1.5 mL | 2×1.5 mL | 2×10 mL |
Applications
1.LAMP isothermal amplificationem
2.DNA filum una obsessio reactionem
3.High GC gene sequencing
4.DNA sequencing de gradu nanogram.
Repono Condition
Transportatio sub 0°C et reponenda -25°C~-15°C.
Unitas Definition
Una unitas definitur quantum enzyme, quod 25 nmol de dNTP in acidum insolubilem materiae 30 minutarum 65°C incorporet.
Regimen quālitātis
1.Dapibus Puritas Asssay (SDS-PAGE):Puritas Bst DNA polymerasis V2 ≥99% determinatur per analysim SDS-Paginae per detectionem Coomassie Blue.
2.Exonuclease Actio:Incubatio 50 μL reactionis continens minimum 8 U Bst DNA polymerasis V2 cum 1 μg λ -Hind digestum DNA per 16 horas ante 37 eventum in nullo detectabili degradationis modo determinato.
3.Nickase Activity:Incubatio 50 μL reactionis continens minimum 8 U Bst DNA polymerasi V2 cum 1 μg pBR322 DNA pro 16 horis ante 37°C eventum nullo degradatione detecta prout determinatum est.
4.RNase Actio:Incubatio 50 μL reactionis continens minimum 8 U Bst DNA polymerasi V2 cum 1.6 μg MS2 RNA per 16 horas ad 37°C eventum nullo detectabili degradationis modo determinatum.
5.E. coli DNA;120 U of Bst DNA polymerase V2 obiectu E. coli genomico DNA adhibito TaqMan qPCR cum primers specificis pro E. coli 16S rRNA locum.The E. coli genomic DNA contagione ≤1 Exemplar est.
LUMEN reactionem
| Components | 25μL |
| 10×HC Bst V2 Buffer | 2.5 μL |
| MgSO4 (100mM) | 1.5 μL |
| dNTPs (10mM quisque) | 3.5 μL |
| SYTO™ 16 Green (25×)a | 1.0 μL |
| Primario mixb | 6 μL |
| Bst DNA Polymerase V2 (Glycerolum liberorum) (8 U/uL) | 1 μL |
| Formula | μL |
| ddH₂O | Usque ad 25 μL |
Notae:
1) a.SYTOTM 16 Sedum (25×): Iuxta necessitates experimentales aliae colores in substitutis adhiberi possunt;
2) b.Prima mix: permixtione 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 et aliis uoluminibus.
Reactio et Condition
1 HC Bst V2 Buffer, temperatura incubationis est inter 60°C et 65°C.
Calor Inactivation
80 °C20mins
