Bilaw diiran 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.Bst DNA polymerase V2 waa nooc bilow ah oo ku salaysan Bst DNA polymerase V2 (HC5005A) oo lagu helay tignoolajiyada wax ka beddelka ah, taas oo joojin karta dhaqdhaqaaqa DNA polymerase heerkulka qolka, markaa habka falcelinta waa lagu shaqayn karaa oo lagu qaabayn karaa heerkulka qolka si looga hortago in aan Kordhinta gaarka ah iyo hagaajinta waxtarka falcelinta, iyo noocaan waa la lyophilized.Intaa waxaa dheer, hawsheeda waxaa lagu sii daayaa heerkul sare, markaa ma jirto baahi loo qabo tallaabo firfircooni gooni ah.
Qaybaha
| Qayb | HC5006A-01 | HC5006A-02 | HC5006A-03 |
Bst DNA polymerase 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.laambad-weynaynta isothermal
2.Xarun DNA oo falcelin barakac ah
3.Taxanaha hiddo-wadaha GC-ga sare
4.Isku xigxiga DNA ee 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.EndonucleaseHawsha:Koritaanka falcelinta 50 μL oo ka kooban ugu yaraan 8 U ee Bst DNA polymerase V2 leh 1 μg DNA 16 saacadood at 37 ℃ natiijadu ma keenayso hoos u dhac la ogaan karo sida la go'aamiyay.
3.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.
4.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.
5.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.
6.E. koliDNA: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
