Panoramica del Prodotto
PCR amplification of DNA fragments, DNA labeling, primer extension, sequence determination, blunt end addition of A, TA vector cloning
| Product Information | |
|---|---|
| Peso molecolare | 94 KD |
| Polymerase Activity | 5′-3′ DNA polimerasi, 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 |
| Sistema di reazione (20ml) | Addition Amount | Remarks |
|---|---|---|
| Pure DNA Polymerase | 0.8ml | Adjust for amplification needs |
| 10x PCR Buffer | 2ml | |
| dNTP (2.5mm) | 2ml | |
| PrimerF | 0.1-1ml | 10-50pmol used |
| PrimerR | 0.1-1ml | 10-50pmol used |
| Template | <1μg | |
| ddH2O | Up to 20μl |
| Cycle Program | Temperatura (°C) | Tempo | Remarks |
|---|---|---|---|
| Initial Denaturation | 94 | 3min | |
| Denaturazione | 94 | 30S (30-40 cicli) | |
| Ricottura | 45-60 | 30S | |
| Estensione | 72 | 30s-2min | Extension time varies with fragment |
| Final Extension | 72 | 10min |
Procedura
Can be directly detected without adding a Loading Buffer
Result Detection
Dopo PCR, Prendere 5 μl of product for agarose gel electrophoresis
Recensioni
Non ci sono ancora recensioni.