Bst 2.0 DNA Polymerase (Glycerol bilaash ah)
Bst DNA polymerase V2 waxaa laga soo qaatay Bacillus stearothermophilus DNA Polymerase I, kaas oo leh 5′→3′ firfircoonida polymerase DNA iyo dhaqdhaqaaqa beddelka silsiladda xooggan, laakiin ma jiro 5′→3′ waxqabadka exonuclease.Bst DNA Polymerase V2 waxay ku habboon tahay barokaca strand-ka, laambad-weynaynta isothermal (loop mediated isothermal amplification) iyo isku xigxig degdeg ah.
Qaybaha
| Qayb | HC5005A-01 | HC5005A-02 | HC5005A-03 |
| BstDNApolymerase V2 (Glycerol-free) (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 |
Codsiyada
1.LAMP isothermal amplification
2.DNA strand hal falcelinta barakaca
3.High GC isku xigxiga
4.DNA isku xigxiga heerka nanogram.
Xaalada Kaydinta
Gaadiidka ka hooseeya 0°C waxaana lagu kaydiyaa -25°C~-15°C.
Qeexitaan Unug
Hal unug ayaa lagu qeexaa cadadka ensaymka ee ku dara 25 nmol ee dNTP walax aan milmin aashitada 30 daqiiqo 65°C.
Xakamaynta tayada
1.Qiimaynta Nadiifinta Barootiinka (SDS-PAGE):Nadiifinta Bst DNA polymerase V2 waa ≥99% waxaa go'aamiyay falanqaynta SDS-PAGE iyadoo la adeegsanayo ogaanshaha Coomassie Blue.
2.Waxqabadka Exonuclease:Koritaanka falcelinta 50 μL oo ka kooban ugu yaraan 8 U ee Bst DNA polymerase V2 oo leh 1 μg λ -Hind Ⅲ dheefshiidka DNA 16 saacadood ee 37 ℃ natiijadu ma keento hoos u dhac la ogaan karo sida la go'aamiyay.
3.Waxqabadka Nickase:Koritaanka falcelinta 50 μL oo ka kooban ugu yaraan 8 U ee Bst DNA polymerase V2 leh 1 μg pBR322 DNA 16 saacadood at 37°C natiijadu ma keenayso hoos u dhac la ogaan karo sida la go'aamiyay.
4.Waxqabadka RNase:Koritaanka falcelinta 50 μL oo ka kooban ugu yaraan 8 U ee Bst DNA polymerase V2 oo leh 1.6 μg MS2 RNA 16 saacadood at 37°C natiijadu ma keenayso hoos u dhac la ogaan karo sida la go'aamiyay.
5.E. coli DNA:120 U of Bst DNA polymerase V2 ayaa laga baadhay joogitaanka E. coli genomic DNA iyadoo la isticmaalayo TaqMan qPCR oo leh qalabyo u gaar ah E. coli 16S rRNA goobta.Faddarada E. coli genomic DNA waa ≤1 Nuqul.
Falcelinta LAMP
| Qaybaha | 25μL |
| 10×HC Bst V2 Buffer | 2.5 μL |
| MgSO4 (100mM) | 1.5 μl |
| dNTPs (10mM midkiiba) | 3.5 μl |
| SYTO™ 16 Cagaar (25×)a | 1.0 μL |
| Isku dhafka koowaadb | 6 μL |
| Bst DNA Polymerase V2 (Glycerol-free) (8 U/ul) | 1 μL |
| Template | × μL |
| ddH₂O | Ilaa 25 μl |
Xusuusin:
1) ah.SYTOTM 16 Cagaaran (25×): Marka loo eego baahida tijaabada ah, midabyo kale ayaa loo isticmaali karaa beddelka;
2) b.Isku dhafka koowaad: waxaa lagu helay isku darka 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 iyo qiyaaso kale.
Falcelinta iyo Xaaladda
1 × HC Bst V2 Buffer, heerkulku wuxuu u dhexeeyaa 60°C iyo 65°C.
Dhaqdhaqaaq la'aanta kulaylka
80 ° C, 20 daqiiqo
