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For PCR amplification of Wolbachia 16S rRNA gene at nucleotides 955–1392.
Forward primer: 5’•CATACCTATTCGAAGGGATAG•3’
Reverse primer: 5’•AGCTTCGAGTGAAACCAATTC•3’
Materials
- Promega GoTaq® Flexi DNA polymerase
- NEB 100 bp ladder
- 1% agarose gel in TBE
Extension temperature is dependent on polymerase
| Step | Temperature (°C) | Time |
| initial denaturation | 94 | 2 min. |
| 30 cycles | 94 | 30 sec. |
| 55 | 45 sec. | |
| 72 | 1 min. | |
| extension | 72 | 10 min. |
| hold | 10 | ∞ |
