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Nipah Virus Nucleoprotein Gene Specific Primers and RT-PCR Conditions
| Primers | ||
| 5′-CTGCTGCAGTTCAGGAAACATCAG-3′ | ||
| 5′-ACCGGATGTGCTCACAGAACTG-3′ | ||
| RT-PCR mix | ||
| Primers (10 pmol/μl) | 5 μl each | |
| dNTP (10 mmol/L) | 4 μl | |
| Buffer (5×) | 10 μl | |
| DTT (100 mmol/L) | 2 μl | |
| RNAse inhibitor (5 U/μl) | 1 μl | |
| Titan enzyme mix | 1 μl | |
| RNA template | 10 to 20 μl | |
| H2O to final volume of 50 μl | ||
| RT-PCR conditions | ||
| 50°C | 30 min | |
| 94°C | 5 min | |
| 94°C | 1 min × 30 cycles | |
| 50°C | 1 min × 30 cycles | |
| 72°C | 2 min × 30 cycles | |
| 72°C | 10 min | |
| 4°C | Hold | |
