Descripción del producto
PCR amplification of DNA fragments, DNA labeling, primer extension, sequence determination, blunt end addition of A, TA vector cloning
| Información del producto | |
|---|---|
| Peso molecular | 94 kd |
| Polymerase Activity | 5′-3′ ADN polimerasa, 5′-3′ exonuclease (no 3′-5′ exonuclease) |
| PCR Buffer | 10 X PCR Buffer for direct loading |
| Experimental Efficiency | The PCR reaction can be directly detected by electrophoresis |
| Extension Speed | 1-2 kb/min |
| Terminal Base | Product has A at the 3′ end, facilitating TA vector cloning |
| Reaction System (20µl) | Addition Amount | Remarks |
|---|---|---|
| Pure DNA Polymerase | 0.8µl | Adjust for amplification needs |
| 10x PCR Buffer | 2µl | |
| dNTP (2.5milímetro) | 2µl | |
| PrimerF | 0.1-1µl | 10-50pmol used |
| PrimerR | 0.1-1µl | 10-50pmol used |
| Template | <1µg | |
| ddH2O | Up to 20μl |
| Cycle Program | Temperatura (°C) | Tiempo | Remarks |
|---|---|---|---|
| Initial Denaturation | 94 | 3mín. | |
| Desnaturalización | 94 | 30s (30-40 ciclos) | |
| Recocido | 45-60 | 30s | |
| Extensión | 72 | 30s-2min | Extension time varies with fragment |
| Final Extension | 72 | 10mín. |
Procedimiento
Can be directly detected without adding a Loading Buffer
Result Detection
Después de la PCR, llevar 5 μl of product for agarose gel electrophoresis
Reseñas
Aún no hay reseñas.