2× PCR Premix (oo leh Midab) HYA442
2× PCR Master Mix wuxuu ka kooban yahay Taq DNA Polymerase, dNTPs, iyo qaybo kale oo PCR looga baahan yahay. Master Mix waa mid deggan muddo 3 bilood ah heerkul ah 4℃ iyadoo la adeegsanayo xasiliyeyaashayada gaarka ah. Xalka isku darka kahor waxaa loo habeeyay PCR-ga caadiga ah wuxuuna diyaar u yahay in la isticmaalo iyadoo lagu darayo shaxanka DNA-da iyo astaamaha hordhaca ah. Badeecadaha PCR waxaa si toos ah loogu shubi karaa electrophoresis iyadoo la isticmaalayo rinji buluug ah oo bromophenol ah oo hore loo sii raray. Badeecadaha la ballaariyay waxay ka kooban yihiin 3 '-dA soo bixitaan waxaana si fudud loogu koobi karaa vector-ka T. 2× PCR Master Mix wuxuu fududeeyaa habka PCR wuxuuna yareeyaa wasakhowga.
Qaybaha
| Qaybaha | Cabbirka-1 | Cabbirka-2 | Cabbirka-3 |
| 2 × PCR Premix (oo leh Midab) | 5mL | 1 mL × 15 | 1 mL × 50 |
Kaydinta
Ku kaydi heerkul ah -20°C muddo 2 sano ah.
Tilmaamaha
1. Nidaamka Falcelinta
| Qaybaha | Mugga |
| 2 × PCR Premix (oo leh Midab) | 25 uL |
| Horudhac Qorrax-dhalaal (10 μM) | 2 µL |
| Ra'iisul-wasaaraha dambe (10 μM) | 2 µL |
| Qaab-dhismeedka DNA* | Doorsoome |
| ddH2THE | Ilaa 50 µL |
* Isticmaalka qaabka: 50-200 ng DNA genomic; 0.1-10 ng DNA plasmid
2. Barnaamijka Falcelinta
| Tallaabo | Heerkulka | Waqtiga | Wareegyo |
| Bilawga Qalitaanka Ilkaha | 94 ℃ | 5 daqiiqo | 1 |
| Denaturation | 94 ℃ | 30 ilbiriqsi | 35 |
| Qaboojinta | 50-60 ℃ | 30 ilbiriqsi | |
| Kordhinta | 72 ℃ | 30-60 ilbiriqsi/kb | |
| Kordhinta Kama Dambeysta ah | 72 ℃ | 10 daqiiqo | 1 |
| Hayso | 4 ℃ | - | 1 |
1. Mg2+ Fiirsashada: Badeecadani waxay ka kooban tahay 3 mM oo MgCl ah2, oo ku habboon inta badan falcelinta PCR.
2. Heerkulka Qaboojinta: Fadlan tixraac qiimaha Tm ee aragtiyeed ee astaamaha. Heerkulka qaboojinta waxaa loo dejin karaa 2-5℃ ka hooseeya qiimaha aragtiyeed ee astaamaha.
3. Waqtiga dheereynta: Si loo aqoonsado molecular-ka, 30 ilbiriqsi/kb ayaa lagu talinayaa. 60 ilbiriqsi/kb ayaa lagu talinayaa.
Qoraallo
1. Badeecadaha PCR ee leh 2× PCR Master Mix kuma habboona electrophoresis jel polyacrylamide.
2. Badbaadadaada iyo caafimaadkaaga awgeed, fadlan xidho jaakadaha shaybaarka iyo galoofyada la tuuri karo si aad u shaqeyso.
3. Waxaa loo isticmaalaa cilmi-baaris oo KALIYA!