I-Wild Taq DNA Polymerase
I-Taq DNA Polymerase iyi-polymerase ye-DNA eshisayo evela ku-Thermus aquaticus YT-1, enomsebenzi we-polymerase ongu-5′→3′ kanye nomsebenzi ongu-5′ flap endonuclease.
Izingxenye
| Isakhi | HC1010A-01 | HC1010A-02 | HC1010A-03 | HC1010A-04 |
| 10× I-Taq Buffer | 2×1 mL | 2 × 10 mL | 2 × 50 mL | 5×200 mL |
| I-Taq DNA Polymerase (5 U/μL) | 0.1 mL | 1 mL | 5 ml | 5 × 10 mL |
Isimo Sesitoreji
Ukuthutha ngaphansi kuka-0°C futhi kugcinwe ku -25°C~-15°C.
Incazelo Yeyunithi
Iyunithi eyodwa ichazwa njengenani le-enzyme elihlanganisa i-15 nmol ye-dNTP kwinto engancibiliki ene-asidi emizuzwini engama-30 ku-75°C.
Ikhwalithi yokulawula
1.I-Protein Purity Assay (SDS-PAGE):Ukuhlanzeka kwe-Taq DNA polymerase kwaba ≥95% kunqunywa ukuhlaziywa kwe-SDS-PAGE.
2.EndUmsebenzi we-onuclease:Ubuncane obungu-5 U ye-Taq DNA polymerase ene-1 μg λDNA amahora angu-16 ku-37 ℃ ibangela ukonakala okubonakalayo njengoba kunqunyiwe.
3.Umsebenzi we-Exonuclease:Ubuncane obungu-5 U ye-Taq DNA polymerase ene-1 μg λ -Hind Ⅲ yokugaya i-DNA amahora angu-16 ngo-37 ℃ iphumela ekubeni kungabikho ukuwohloka okubonakalayo njengoba kunqunyiwe.
4.Umsebenzi we-Nickase:Ubuncane obungu-5 U ye-Taq DNA polymerase ene-1 μg pBR322 DNA amahora angu-16 ku-37°C ibangela ukuwohloka okubonakalayo njengoba kunqunyiwe.
5.Umsebenzi we-RNase:Ubuncane obungu-5 U ye-Taq DNA polymerase ene-1.6 μg MS2 RNA amahora angu-16 ku-37°C ibangela ukonakala okubonakalayo njengoba kunqunyiwe.
6.E. coliI-DNA:I-5 U ye-Taq DNA polymerase ihlolelwa ukuba khona kwe-E. coli genomic DNA kusetshenziswa i-TaqMan qPCR eneziqalo eziqondile ze-E. coli 16S rRNA locus.Ukungcola kwe-E. coli genomic DNA kungu-≤1 Ikhophi.
7.I-PCR Amplification (5.0 kb Lambda DNA)- Ukusabela okungu-50 µL okuqukethe i-5 ng Lambda DNA enamayunithi angu-5 e-Taq DNA Polymerase yemijikelezo engu-25 yemiphumela yokukhulisa i-PCR kumkhiqizo olindelwe ongu-5.0 kb.
Ukusethwa kokusabela
| Izingxenye | Ivolumu |
| I-DNA yesifanekisoa | ngokuzikhethela |
| 10 μM Phambili I-Primer | 1 μl |
| 10 μM I-Primer Reverse | 1 μl |
| I-dNTP Mix (10mM ngayinye) | 1 μl |
| 10×I-Taq Buffer | 5 μl |
| I-Taq DNA Polymeraseb | 0.25 μL |
| Amanzi angenayo i-nuclease | Kufika ku-50 μL |
Amanothi:
1) I-concentration yokusabela efanele yezifanekiso ezahlukene ihlukile.Ithebula elilandelayo libonisa ukusetshenziswa kwesifanekiso esinconyiwe sesistimu yokusabela engu-50 µL.
| I-DNA | Inani |
| I-Genomic | 1 ng-1 μg |
| I-Plasmid noma i-Viral | 1 ikhasi-1 ng |
2) Ukugxiliswa okuphelele kwe-Taq DNA Polymerase kungase kusuke ku-0.25 µL~1 µL ezinhlelweni ezikhethekile.
UkusabelaUhlelo
| Isinyathelo | Izinga lokushisa(°C) | Isikhathi | Imijikelezo |
| I-denaturation yokuqalaa | 95 ℃ | 5 imiz | - |
| I-Denaturation | 95 ℃ | 15-30 s | 30-35 Imijikelezo |
| Anealingb | 60 ℃ | 15 ik | |
| Isandiso | 72 ℃ | 1kb/min | |
| Isandiso Sokugcina | 72 ℃ | 5 imiz | - |
Amanothi:
1) Isimo sokuqala se-denaturation sifanele ukusabela okuningi kokukhulisa futhi singashintshwa ngokuya ngobunkimbinkimbi besakhiwo sesifanekiso.Uma isakhiwo sesifanekiso siyinkimbinkimbi, isikhathi sangaphambi kokuguqula singanwetshwa sibe yi-5 - 10mins ukuze kuthuthukiswe umphumela wokuqala wokuguqula.
2) Izinga lokushisa le-anneal lidinga ukulungiswa ngokuvumelana nenani le-Tm le-primer, ngokuvamile elibekwe ku-3~5 ℃ ngaphansi kwevelu ye-Tm ye-primer;Kuzifanekiso eziyinkimbinkimbi, kuyadingeka ukulungisa izinga lokushisa le-anneal nokwelula isikhathi sokunwetshwa ukuze kuzuzwe ukukhuliswa okuphumelelayo.
