Duur Taq DNA Polymerase
Taq DNA Polymerase waa kuleyl DNA polymerase ah oo ka yimaada Thermus aquaticus YT-1, kaas oo leh 5′→3′ firfircoonida polymerase iyo 5′ flap endonuclease act.
Qaybaha
| Qayb | HC1010A-01 | HC1010A-02 | HC1010A-03 | HC1010A-04 |
| 10× Taq Buffer | 2×1 ml | 2×10 ml | 2×50 ml | 5×200 ml |
| Taq DNA Polymerase (5 U/μL) | 0.1 ml | 1 ml | 5 ml | 5×10 ml |
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.Qiimaynta Nadiifinta Barootiinka (SDS-PAGE):Nadiifinta Taq DNA polymerase waxaa ≥95% lagu go'aamiyay falanqaynta SDS-PAGE.
2.EndWaxqabadka onuclease:Ugu yaraan 5 U of Taq DNA polymerase leh 1 μg λDNA 16 saacadood at 37 ℃ natiijadu ma aha hoos u dhac la ogaan karo sida la go'aamiyay.
3.Waxqabadka Exonuclease:Ugu yaraan 5 U of Taq DNA polymerase oo leh 1 μg λ -Hind Ⅲ dheefshiidka DNA 16 saacadood at 37 ℃ natiijadu ma aha hoos u dhac la ogaan karo sida la go'aamiyay.
4.Waxqabadka Nickase:Ugu yaraan 5 U of Taq DNA polymerase leh 1 μg pBR322 DNA 16 saacadood at 37°C natiijadu ma jirto hoos u dhac la ogaan karo sida la go'aamiyay.
5.Waxqabadka RNase:Ugu yaraan 5 U ee Taq DNA polymerase oo leh 1.6 μg MS2 RNA 16 saacadood oo ah 37°C natiijadu ma jirto wax hoos u dhac la ogaan karo sida la go'aamiyay.
6.E. koliDNA:5 U of Taq DNA polymerase waxaa lagu baara 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.
7.Kordhinta PCR (5.0kb Lambda DNA)- Falcelinta 50 µL oo ka kooban 5 ng Lambda DNA oo leh 5 unug Taq DNA Polymerase ee 25 wareegyada natiijooyinka kordhinta PCR ee badeecada 5.0kb ee la filayo.
Dejinta falcelinta
| Qaybaha | Mugga |
| Qaabka DNA-daa | ikhtiyaari ah |
| 10 μM Horudhac Hore | 1 μL |
| 10 μM Reverse Primer | 1 μL |
| Isku-dhafka dNTP (10mM midkiiba) | 1 μL |
| 10× Taq Buffer | 5 μL |
| Taq DNA Polymeraseb | 0.25 μl |
| Biyo aan nukliyeer lahayn | Ilaa 50 μl |
Xusuusin:
1) Xoojinta falcelinta ugu fican ee qaabab kala duwan ayaa ka duwan.Jadwalka soo socdaa wuxuu muujinayaa qaabka lagu taliyey ee isticmaalka 50 µL habka falcelinta.
| DNA | Qadarka |
| Genomic | 1 ng-1 μg |
| Plasmid ama Viral | 1 pg-1 ng |
2) U-fiirsashada ugu fiican ee Taq DNA Polymerase waxay u dhaxayn kartaa 0.25 µL ~ 1 µL codsiyada khaaska ah.
FalcelintaBarnaamijka
| Talaabada | Heerkulka(°C) | Waqtiga | Wareegtada |
| Denaturation bilowga aha | 95 ℃ | 5 daqiiqo | - |
| Denaturation | 95 ℃ | 15-30 s | 30-35 wareeg |
| Annealingb | 60 ℃ | 15 s | |
| Kordhinta | 72 ℃ | 1kb/min | |
| Kordhinta u dambaysa | 72 ℃ | 5 daqiiqo | - |
Xusuusin:
1) Xaaladda denaturation bilowga ah waxay ku habboon tahay inta badan falcelinta xoojinta waxaana wax laga beddeli karaa iyadoo loo eegayo kakanaanta qaab-dhismeedka template.Haddii qaab-dhismeedka qaab-dhismeedka uu yahay mid adag, wakhtiga ka-hor-istaagga waxaa la kordhin karaa 5 - 10mins si loo hagaajiyo saamaynta denaturation bilowga ah.
2) Heerkulka xajinta wuxuu u baahan yahay in lagu hagaajiyo iyadoo loo eegayo qiimaha Tm ee asaasiga ah, kaas oo guud ahaan lagu dejiyay 3 ~ 5 ℃ ka hooseeya qiimaha Tm ee asaasiga ah;Qaababka kakan, waxaa lagama maarmaan ah in la hagaajiyo heerkulka jilbiska iyo kordhinta waqtiga fidinta si loo gaaro kor u qaadis hufan.
