Bst 2.0 DNA Polymerase (Glycerol yaulere, yochuluka kwambiri)
Bst DNA polymerase V2 imachokera ku Bacillus stearothermophilus DNA Polymerase I, yomwe ili ndi 5'→ 3' DNA polymerase ntchito ndi ntchito yamphamvu yowonjezera unyolo, koma palibe 5′→3′ exonuclease ntchito.Bst DNA Polymerase V2 ndiyoyenera kusamuka kwa strand, isothermal amplification LAMP (Loop mediated isothermal amplification) ndikutsatizana mwachangu.Bst DNA polymerase V2 iyi imatha kuletsa ntchito ya DNA polymerase kutentha kwa firiji, kuti igwire ntchito komanso momwe zimachitikira zimatha kupangidwa ndi kutentha kwa firiji, kuteteza kukulitsa kosakhazikika komanso kuwongolera magwiridwe antchito, ndipo mtundu uwu ukhoza kukhala lyophilized.Kuonjezera apo, imatha kumasula ntchito yake pa kutentha kwakukulu, motero kuchotsa kufunikira kwa sitepe yapadera yoyambitsa.
Zigawo
| Chigawo | Mtengo wa HC5007A-01 | Mtengo wa HC5007A-02 | Mtengo wa HC5007A-03 |
| Bst DNA polymerase V2 (Glycerol-free) (32U/μL) | 0.05 ml ya | 0.25 mL | 2.5 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 |
Mapulogalamu
1.LAMP isothermal amplification
2.DNA strand single displacement reaction
3.High GC gene kutsatizana
4.Kutsata kwa DNA kwa nanogram level.
Mkhalidwe Wosungira
Kuyenda pansi pa 0 ° C ndikusungidwa pa -25 ° C ~ -15 ° C.
Tanthauzo la Chigawo
Chigawo chimodzi chimatanthauzidwa ngati kuchuluka kwa enzyme yomwe imaphatikiza 25 nmol ya dNTP muzinthu zosasungunuka za asidi mu mphindi 30 pa 65 ° C.
Kusintha kwa LAMP
| Zigawo | 25 μLDongosolo |
| 10 × HC Bst V2 Buffer | 2.5 μL |
| MgSO4 (100mM) | 1.5 μL |
| dNTPs (10mM iliyonse) | 3.5 μl |
| SYTO™ 16 Green (25×)a | 1.0 μL |
| Kusakaniza koyambirirab | 6 ml |
| Bst DNA Polymerase V2 (Glycerol-free) (32 U/uL) | 0.25 μL |
| Template | × μl ndi |
| ddH₂O | Mpaka 25 μL |
Ndemanga:
1) a.SYTOTM 16 Green (25 ×): Malingana ndi zosowa zoyesera, utoto wina ukhoza kugwiritsidwa ntchito m'malo;
2) b.Kusakaniza koyamba: kupezedwa posakaniza 20 µM FIP, 20 µM BIP, 2.5 µM F3, 2.5 µM B3, 5 µM LF, 5 µM LB ndi mavoliyumu ena.
Zochita ndi Mkhalidwe
1 × HC Bst V2 Buffer, kutentha kwa ma incubation kuli pakati pa 60°C ndi 65°C.
Kutentha Kutentha
80 ° C, mphindi 20.
