|
| ||
| Component | Amount (μl) | Final concentration |
|
| ||
| Q5 Reaction Buffer (5×) | 10 | 1× |
|
| ||
| dNTPs (10 mM each) | 1 | 0.2 mM each |
|
| ||
| UniFwd primer (25 μM) | 1 | 0.5 μM |
|
| ||
| UniRev primer (25 μM) | 1 | 0.5 μM |
|
| ||
| Purified Step 1 PCR product | 10 | |
|
| ||
| Q5 Hot-Start DNA Polymerase (2 U/μl) | 0.5 | 0.02 U/μl |
|
| ||
| Nuclease-free water | 26.5 | |
|
| ||
| Final | 50 (for one reaction) | |
|
| ||
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