Inodziya Kutanga 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.Bst DNA polymerase V2 ndiyo inopisa-yekutanga vhezheni yakavakirwa paBst DNA polymerase V2 (HC5005A) inowanikwa neinodzoreredza gadziriso tekinoroji, iyo inogona kutadzisa DNA polymerase chiitiko pakupisa kwekamuri, saka iyo reaction system inogona kushandiswa uye kuumbwa pakamuri tembiricha kudzivirira kusavapo. -specific amplification uye kuvandudza maitiro ekuita, uye iyi vhezheni inogona kuve lyophilized.Mukuwedzera, basa rayo rinosunungurwa pakupisa kwepamusoro, saka hapana chikonzero chekugadzirisa danho rakasiyana.
Zvikamu
| Chikamu | HC5006A-01 | HC5006A-02 | HC5006A-03 |
Bst DNA polymerase 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.Yakakwira 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.EndonucleaseChiitiko:Kuiswa kwe50 μL reaction ine hushoma hwe8 U yeBst DNA polymerase V2 ine 1 μg λDNA kwemaawa gumi nematanhatu pa37 ℃ inokonzeresa kushatisa kunoonekwa sekurongwa.
3.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.
4.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.
5.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.
6.E. coliDNA: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
