Hotstart Taq DNA Polymerase
Hot Start Taq DNA Polymerase (Beddelka Antibody) waa kulayl-bilawga DNA polymerase ka Thermus aquaticus YT-1, kaas oo leh 5′→3′ firfircoonida polymerase iyo 5′ flap endonuclease act.Bilawga kulul ee Taq DNA polymerase waa Taq DNA polymerase kaas oo ay wax ka beddeleen unugyada difaaca jirka ee Taq.Wax ka beddelka ka-hortagga jidhka ayaa kordhay gaar ahaan, dareenka, iyo dhalidda PCR.
Qaybaha
| Qayb | HC1012A-01 | HC1012A-02 | HC1012A-03 | HC1012A-04 |
| 5×HC Taq Buffer | 4×1 ml | 4×10 ml | 4×50 ml | 5×400 ml |
| Bilawga Kulul ee Taq DNA Polymerase (Antibody modified) (5 U/μL) | 0.1 ml | 1 ml | 5 ml | 10×5 mL |
Codsiyada
10mM Tris-HCl (pH 7.4 at 25℃), 100 mM KCl, 0.1 mM EDTA, 1 mM dithiothreitol, 0.5% Tween20, 0.5% IGEPALCA-630 iyo 50% Glycerol.
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 15 nmol ee dNTP walax aan milmin aashitada 30 daqiiqo 75°C.
Xakamaynta tayada
1.EndWaxqabadka onuclease:Ku-soo-baxa 20 U of enzyme leh 4 μg pUC19 DNA 4 saacadood oo ah 37 ℃ waxay keentay in aan la ogaan karin hoos u dhac ku yimaada DNA-da sida lagu go'aamiyay jel electrophoresis.
2.5 kb Lambda PCR:25 wareegyada PCR ee 5 ng Lambda DNA oo leh 1.25 unug Taq DNA Polymerase iyadoo ay joogaan 200 µM dNTPs iyo 0.2 µM natiijooyinka 5 kb ee la filayo.
3.Waxqabadka Exonuclease:Kordhinta falcelinta 50 µl oo ka kooban ugu yaraan 12.5 U of Taq DNA Polymerase leh 10 nmol 5′-FAM oligonucleotide 30 daqiiqo 37℃ ma keento hoos u dhac la ogaan karo.
4.Waxqabadka RNase:Kordhinta falcelinta 10 µL oo ka kooban 20 U of enzyme leh 1μg ee qoraallada RNA muddo 2 saacadood ah 37°C waxay keentay in aan la ogaan karin hoos u dhac RNA ah sida lagu go'aamiyay jel electrophoresis.
5.Dhaqdhaqaaq la'aanta kulaylka:Maya
Nidaamka falcelinta
| Qaybaha | Mugga |
| Qaabka DNA-daa | ikhtiyaari ah |
| 10 μM Horudhac Hore | 0.5 μl |
| 10 μM Reverse Primer | 0.5 μl |
| Isku-dhafka dNTP (10mM midkiiba) | 0.5 μl |
| 5×HC Taq Buffer | 5 μL |
| Taq DNA Polymeraseb(5U/μL) | 0.125 μl |
| Biyo aan nukliyeer lahayn | Ilaa 25 μl |
Xusuusin:
1) ah.
| DNA | Qadarka |
| Genomic | 1 ng-1 μg |
| Plasmid ama Viral | 1 pg-1 ng |
2) b.U fiirsashada ugu fiican ee Taq DNA Polymerase waxay u dhaxaysaa 5-50 unug/ml (0.1-0.5 unug/25 µL falcelinta) ee codsiyada gaarka ah.
Nidaamka baaskiil wadida kulaylka
PCR
| Talaabada | Heerkulka(°C) | Waqtiga | Wareegtada |
| Denaturation bilowga aha | 95 ℃ | 1-3 daqiiqo | - |
| Denaturation | 95 ℃ | 15-30 s | 30-35 wareeg |
| Annealingb | 45-68 ℃ | 15-60 s | |
| Kordhinta | 68 ℃ | 1kb/min | |
| Kordhinta u dambaysa | 68 ℃ | 5 daqiiqo | - |
Xusuusin:
1) Denaturation bilowga ah ee 1 daqiiqo at 95°C ayaa ku filan badibaQaababka adag, diidmo dheer oo ah 2-3mins at 95°C ayaa lagula talinayaa.PCR-da colony, 5mins denaturation oo ah 95°C ayaa lagu talinayaa.
2) Talaabada annealing caadi ahaan waa 15-60 s.Heerkulka xinjirtu wuxuu ku salaysan yahay Tm ee lammaanaha asaasiga ah oo caadi ahaan waa 45-68 ℃.
