Bst 2.0 DNA Polymerase (Glycerol yemahara)
Bst DNA polymerase V2 inotorwa kubva kuBacillus stearothermophilus DNA Polymerase I, iyo ine 5′→ 3′ DNA polymerase basa uye yakasimba cheni kutsiva basa, asi hapana 5′→3′ exonuclease basa.Bst DNA Polymerase V2 yakanyatsokodzera strand-displacement, isothermal amplification LAMP (Loop mediated isothermal amplification) uye kukurumidza kutevedzana.
Zvikamu
| Chikamu | HC5005A-01 | HC5005A-02 | HC5005A-03 |
| BstDNApolymerase V2 (Glycerol-isina) (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 amplification
2.DNA strand single displacement reaction
3.High GC gene sequencing
4.DNA sequencing ye nanogram level.
Storage Condition
Kutakura pasi pe 0°C uye kuchengetwa pa -25°C~-15°C.
Chikamu Tsanangudzo
Chimwe chikamu chinotsanangurwa sehuwandu hwe enzyme inobatanidza 25 nmol ye dNTP mu acid isinganyunguriki zvinhu mumaminitsi makumi matatu pa 65 ° C.
Quality Control
1.Protein Purity Assay (SDS-PEJI):Kuchena kweBst DNA polymerase V2 ndeye ≥99% inotemerwa neSDS-PAGE ongororo uchishandisa Coomassie Blue kuona.
2.Exonuclease Chiitiko:Kuiswa kwe50 μL reaction ine hushoma hwe8 U yeBst DNA polymerase V2 ine 1 μg λ -Hind Ⅲ digest DNA kwemaawa gumi nematanhatu pa37 ℃ inokonzeresa kusadzikiswa kunooneka sekutsanangurwa.
3.Nickase Chiitiko:Kuiswa kwe50 μL reaction ine hushoma hwe8 U yeBst DNA polymerase V2 ine 1 μg pBR322 DNA kwemaawa gumi nematanhatu pa37°C inokonzeresa kushatisa kunoonekwa sekurongwa.
4.RNase Chiitiko:Kuiswa kwe50 μL reaction ine hushoma hwe8 U yeBst DNA polymerase V2 ine 1.6 μg MS2 RNA kwemaawa gumi nematanhatu pa37°C inokonzeresa kushatisa kunoonekwa sekutsanangurwa kwazvakaitwa.
5.E. coli DNA:120 U yeBst DNA polymerase V2 inoongororwa kuvapo kweE. coli genomic DNA uchishandisa TaqMan qPCR ine maprimers chaiwo eE. coli 16S rRNA locus.Iyo E. coli genomic DNA kusvibiswa ndeye ≤1 Copy.
LAMP Reaction
| Zvikamu | 25μL |
| 10 × HC Bst V2 Buffer | 2.5 μL |
| MgSO4 (100mM) | 1.5 μL |
| dNTPs (10mM imwe neimwe) | 3.5 μL |
| SYTO™ 16 Girini (25×)a | 1.0 μL |
| Primer mixb | 6 μL |
| Bst DNA Polymerase V2 (Glycerol-isina) (8 U/uL) | 1 μL |
| Template | × μL |
| ddH₂O | Kusvika ku25 μL |
Notes:
1) a.SYTOTM 16 Green (25 ×): Maererano nezvinodiwa zvekuedza, mamwe madhayi anogona kushandiswa seanotsiva;
2) b.Primer mix: inowanikwa nekusanganisa 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 nemamwe mavhoriyamu.
Kuita uye Mamiriro ezvinhu
1 × HC Bst V2 Buffer, incubation tembiricha iri pakati pe60°C ne65°C.
Heat Inactivation
80 °C, 20mins
