Bst 2.0 ADN Polymerase (Glycerol yubusa)
Bst ADN polymerase V2 yakomotse kuri Bacillus stearothermophilus ADN Polymerase I, ifite 5 ′ → 3 ′ ibikorwa bya polymerase ya ADN nigikorwa gikomeye cyo gusimbuza urunigi, ariko nta gikorwa cya 5 ′ → 3 ′.Bst ADN Polymerase V2 nibyiza cyane muburyo bwo kwimura imirongo, kwimura isothermal amplification LAMP (Loop mediated isothermal amplification) hamwe nuburyo bwihuse.
Ibigize
| Ibigize | HC5005A-01 | HC5005A-02 | HC5005A-03 |
| BstDNApolymerase V2 (Glycerol-yubusa) (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 |
Porogaramu
1.LAMP isothermal amplification
2.DNA umurongo umwe wimurwa
3.Icyiciro cya GC gikurikiranye
4.DNA ikurikirana urwego rwa nanogram.
Imiterere y'Ububiko
Ubwikorezi buri munsi ya 0 ° C kandi bubikwa kuri -25 ° C ~ -15 ° C.
Igisobanuro
Igice kimwe gisobanurwa nkubunini bwa enzyme yinjiza nmol 25 ya dNTP mubintu bitangirika aside muminota 30 kuri 65 ° C.
Kugenzura ubuziranenge
1.Isuzuma rya poroteyine (SDS-PAGE):Ubuziranenge bwa Bst ADN polymerase V2 ni ≥99% bigenwa nisesengura rya SDS-PAGE ukoresheje Coomassie Blue detection.
2.Exonuclease Igikorwa:Inkubasi ya 50 μL reaction irimo byibura 8 U ya Bst ADN polymerase V2 hamwe na 1 μg λ -Hind Ⅲ igogora ADN amasaha 16 kuri 37 ℃ bivamo nta kwangirika kugaragara nkuko byagenwe.
3.Igikorwa cya Nickase:Inkubasi ya 50 μL reaction irimo byibura 8 U ya Bst ADN polymerase V2 hamwe na 1 μg pBR322 ADN kumasaha 16 kuri 37 ° C bivamo nta kwangirika kugaragara nkuko byagenwe.
4.Igikorwa cya RNase:Inkubasi ya 50 μL reaction irimo byibura 8 U ya Bst ADN polymerase V2 hamwe na 1,6 μg MS2 RNA kumasaha 16 kuri 37 ° C bivamo nta kwangirika kugaragara nkuko byagenwe.
5.E. coli ADN:120 U ya Bst ADN polymerase V2 isuzumwa kugirango ADN genomic E. coli ikoreshe TaqMan qPCR hamwe na primers yihariye ya E. coli 16S rRNA.E. coli genomic ADN yanduye ni ≤1 Gukoporora.
Amatara
| Ibigize | 25μL |
| 10 × HC Bst V2 Buffer | 2.5 μL |
| MgSO4 (100mM) | 1.5 μL |
| DNTPs (10mM buri umwe) | 3.5 μL |
| SYTO ™ 16 Icyatsi (25 ×)a | 1.0 μL |
| Kuvanga primerb | 6 μL |
| Bst ADN Polymerase V2 (Glycerol idafite) (8 U / uL) | 1 μL |
| Inyandikorugero | × μL |
| ddH₂O | Kugera kuri 25 μL |
Inyandiko:
1) a.SYTOTM 16 Icyatsi (25 ×): Ukurikije ubushakashatsi bukenewe, andi marangi arashobora gukoreshwa nkibisimburwa;
2) b.Kuvanga primer: byabonetse mukuvanga 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 nibindi bitabo.
Imyitwarire n'imiterere
1 × HC Bst V2 Buffer, ubushyuhe bwa incubation buri hagati ya 60 ° C na 65 ° C.
Gushyushya
80 ° C ， 20min
