Bst 2.0 DNA Polymerase(Glycerol ọfẹ)
Bst DNA polymerase V2 wa lati Bacillus stearothermophilus DNA Polymerase I, eyiti o ni iṣẹ 5′→3′ DNA polymerase ati iṣẹ rirọpo pq ti o lagbara, ṣugbọn ko si iṣẹ-ṣiṣe 5′→3′ exonuclease.Bst DNA Polymerase V2 jẹ apere fun gbigbe-nipo okun, isothermal ampilifaya LAMP (Afikun isothermal mediated Loop) ati itọsẹ kiakia.
Awọn eroja
| Ẹya ara ẹrọ | HC5005A-01 | HC5005A-02 | HC5005A-03 |
| BstDNApolymerase V2(Glycerol-ọfẹ)(8U/μL) | 0.2 milimita | 1 milimita | 10 milimita |
| 10×HC Bst V2 saarin | 1,5 milimita | 2×1.5 milimita | 3×10 milimita |
| MgSO4(100mM) | 1,5 milimita | 2×1.5 milimita | 2×10 milimita |
Awọn ohun elo
1.LAMP isothermal ampilifaya
2.DNA okun nikan nipo nipo lenu
3.High GC jiini lesese
Ilana 4.DNA ti ipele nanogram.
Ibi ipamọ Ipo
Gbigbe labẹ 0°C ati ki o wa ni ipamọ ni -25°C~-15°C.
Unit Definition
Ẹyọ kan jẹ asọye bi iye henensiamu ti o ṣafikun 25 nmol ti dNTP sinu ohun elo airotẹlẹ acid ni ọgbọn iṣẹju ni 65°C.
Iṣakoso didara
1.Ayẹwo Iwa Amuaradagba (SDS-PAGE):Iwa-mimọ ti Bst DNA polymerase V2 jẹ ≥99% ti pinnu nipasẹ itupalẹ SDS-PAGE nipa lilo wiwa Coomassie Blue.
2.Iṣẹ-ṣiṣe Exonuclease:Imudaniloju iṣesi 50 μL ti o ni o kere ju 8 U ti Bst DNA polymerase V2 pẹlu 1 μg λ -Hind Ⅲ digest DNA fun awọn wakati 16 ni awọn abajade 37 ℃ ni ko si ibajẹ wiwa bi a ti pinnu.
3.Nickase aṣayan iṣẹ-ṣiṣe:Iṣeduro iṣesi 50 μL ti o ni o kere ju 8 U ti Bst DNA polymerase V2 pẹlu 1 μg pBR322 DNA fun awọn wakati 16 ni 37°C awọn abajade ni ko si ibajẹ ti o rii bi a ti pinnu.
4.Iṣẹ RNase:Iṣeduro iṣesi 50 μL ti o ni o kere ju 8 U ti Bst DNA polymerase V2 pẹlu 1.6 μg MS2 RNA fun awọn wakati 16 ni awọn abajade 37°C ni ko si ibajẹ wiwa bi a ti pinnu.
5.E. coli DNA:120 U ti Bst DNA polymerase V2 jẹ iboju fun wiwa E. coli genomic DNA nipa lilo TaqMan qPCR pẹlu awọn alakoko kan pato fun E. coli 16S rRNA agbegbe.E. coli jenomic DNA kontaminesonu jẹ ≤1 Daakọ.
Atupa lenu
| Awọn eroja | 25μL |
| 10×HC Bst V2 saarin | 2.5 μL |
| MgSO4 (100mM) | 1.5 μL |
| dNTPs (10mM kọọkan) | 3.5 μL |
| SYTO™ 16 Alawọ ewe (25×)a | 1.0 μL |
| Alakoko illab | 6 μL |
| Bst DNA Polymerase V2 (Glycerol-ọfẹ) (8 U/ul) | 1 μL |
| Àdàkọ | × μL |
| ddH₂O | Titi di 25 μl |
Awọn akọsilẹ:
1) a.SYTOTM 16 Green (25×): Gẹgẹbi awọn iwulo idanwo, awọn awọ miiran le ṣee lo bi aropo;
2) b.Iparapọ alakoko: ti a gba nipasẹ didapọ 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 ati awọn ipele miiran.
Ifesi ati ipo
1 × HC Bst V2 Buffer, iwọn otutu idawọle wa laarin 60°C ati 65°C.
Ooru Inactivation
80 °C, 20 iṣẹju
