Produktübersicht
PCR amplification of DNA fragments, DNA-Markierung, primer extension, sequence determination, blunt end addition of A, TA vector cloning
| Produktinformationen | |
|---|---|
| Molekulargewicht | 94 KD |
| Polymerase Activity | 5′-3′ DNA -Polymerase, 5′-3′ exonuclease (no 3′-5′ exonuclease) |
| PCR-Puffer | 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′ Ende, facilitating TA vector cloning |
| Reaktionssystem (20μl) | Addition Amount | Remarks |
|---|---|---|
| Pure DNA Polymerase | 0.8μl | Adjust for amplification needs |
| 10x PCR Buffer | 2μl | |
| Dntp (2.5mm) | 2μl | |
| PrimerF | 0.1-1μl | 10-50pmol used |
| PrimerR | 0.1-1μl | 10-50pmol used |
| Vorlage | <1μg | |
| ddh2o | Up to 20μl |
| Cycle Program | Temperatur (°C) | Zeit | Remarks |
|---|---|---|---|
| Initial Denaturation | 94 | 3Mindest | |
| Denaturierung | 94 | 30S (30-40 Fahrräder) | |
| Glühen | 45-60 | 30S | |
| Verlängerung | 72 | 30s-2min | Extension time varies with fragment |
| Final Extension | 72 | 10Mindest |
Verfahren
Can be directly detected without adding a Loading Buffer
Result Detection
Nach PCR, nehmen 5 μl of product for agarose gel electrophoresis
Rezensionen
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