Bst 2.0 DNA Polymerase (Glycerol dawb)
Bst DNA polymerase V2 yog muab los ntawm Bacillus stearothermophilus DNA Polymerase I, uas muaj 5′ → 3′ DNA polymerase kev ua thiab muaj zog saw hloov ua haujlwm, tab sis tsis muaj 5′→ 3′ exonuclease kev ua.Bst DNA Polymerase V2 yog qhov tsim nyog rau strand-hloov chaw, isothermal amplification LAMP (Loop mediated isothermal amplification) thiab ceev sequencing.
Cheebtsam
| Cheebtsam | HC5005A-01 | HC5005A-02 | HC5005A-03 |
| BstDNApolymerase V2 (Glycerol-dawb) (8U / μL) | 0.2 ml ib | 1 ml ib | 10 ml ib |
| 10 × HC Bst V2 Buffer | 1.5 ml yog ' | 2 × 1.5 mL | 3 × 10 ml |
| MgSO4(100 hli) | 1.5 ml yog ' | 2 × 1.5 mL | 2 × 10 ml |
Daim ntawv thov
1.LAMP isothermal amplification
2.DNA strand ib qho kev hloov pauv hloov
3.High GC gene sequencing
4.DNA sequencing ntawm nanogram qib.
Kev cia khoom
Kev thauj mus los hauv qab 0 ° C thiab khaws cia ntawm -25 ° C ~ -15 ° C.
Unit txhais
Ib chav tsev tau txhais tias yog tus nqi ntawm cov enzyme uas suav nrog 25 nmol ntawm dNTP rau hauv cov khoom siv acid insoluble hauv 30 feeb ntawm 65 ° C.
Kev Tswj Xyuas Zoo
1.Protein Purity Assay (SDS-PAGE):Lub purity ntawm Bst DNA polymerase V2 yog ≥99% txiav txim los ntawm SDS-PAGE tsom xam siv Coomassie Blue nrhiav.
2.Kev Ua Haujlwm Exonuclease:Incubation ntawm 50 μL cov tshuaj tiv thaiv uas muaj qhov tsawg kawg nkaus ntawm 8 U ntawm Bst DNA polymerase V2 nrog 1 μg λ -Hind Ⅲ zom DNA rau 16 teev ntawm 37 ℃ ua rau tsis pom qhov degradation raws li kev txiav txim.
3.Nkaum kev ua si:Incubation ntawm 50 μL cov tshuaj tiv thaiv uas muaj qhov tsawg kawg nkaus ntawm 8 U ntawm Bst DNA polymerase V2 nrog 1 μg pBR322 DNA rau 16 teev ntawm 37 ° C ua rau tsis pom qhov degradation raws li kev txiav txim.
4.RNase Kev Ua Haujlwm:Incubation ntawm 50 μL cov tshuaj tiv thaiv uas muaj qhov tsawg kawg nkaus ntawm 8 U ntawm Bst DNA polymerase V2 nrog 1.6 μg MS2 RNA rau 16 teev ntawm 37 ° C ua rau tsis pom qhov degradation raws li kev txiav txim.
5.E. coli DNA:120 U ntawm Bst DNA polymerase V2 yog tshuaj ntsuam xyuas qhov muaj E. coli genomic DNA siv TaqMan qPCR nrog primers tshwj xeeb rau E. coli 16S rRNA locus.E. coli genomic DNA paug yog ≤1 Daim.
LAMP Reaction
| Cheebtsam | 25μL |
| 10 × HC Bst V2 Buffer | 2.5 l ua |
| MgSO4 (100 hli) | 1.5 l ua |
| dNTPs (10mM txhua) | 3,5l ua |
| SYTO™ 16 Green (25 ×)a | 1.0 l os |
| primer mixb | 6l miv |
| Bst DNA Polymerase V2 (Glycerol-dawb) (8 U/uL) | 1 mL ib |
| Template | × μL |
| ddH₂O | Txog li 25 μL |
Nco tseg:
1) a.SYTOTM 16 Green (25 ×): Raws li kev sim xav tau, lwm yam xim tuaj yeem siv los hloov pauv;
2) ib.Primer mix: tau los ntawm kev sib xyaw 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 thiab lwm yam ntim.
Cov tshuaj tiv thaiv thiab mob
1 × HC Bst V2 Buffer, qhov ntsuas kub ntawm 60 ° C thiab 65 ° C.
Kub Inactivation
80 ° C, 20 mins
