Product Overview
PCR amplification of DNA fragments, DNA labeling, primer extension, sequence determination, blunt end addition of A, TA vector cloning
| Product Information | |
|---|---|
| Molecular Weight | 94 KD |
| Polymerase Activity | 5′-3′ DNA polymerase, 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 (2.5mM) | 2μl | |
| PrimerF | 0.1-1μl | 10-50pmol used |
| PrimerR | 0.1-1μl | 10-50pmol used |
| Template | <1μg | |
| ddH2O | Up to 20μl |
| Cycle Program | Temperature (°C) | Time | Remarks |
|---|---|---|---|
| Initial Denaturation | 94 | 3min | |
| Denaturation | 94 | 30s (30-40 cycles) | |
| Annealing | 45-60 | 30s | |
| Extension | 72 | 30s-2min | Extension time varies with fragment |
| Final Extension | 72 | 10min |
Procedure
Can be directly detected without adding a Loading Buffer
Result Detection
After PCR, take 5 μl of product for agarose gel electrophoresis
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