Difference between revisions of "FosCreER DREADD PCR"
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(Created page with "Protocol for extraction and purification of DNA from FosCreER x DREADD transgenic colony Category:Protocols") |
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| + | =Extraction and Purification of DNA from Tail Biopsy= | ||
| + | '''Tail Biopsy''' | ||
| + | Mice are first anesthetized in a bell jar with Isoflurane for ~20 sec, monitor animal closely and check with toe pinch before proceeding | ||
| + | |||
| + | =Cycling Protocol= | ||
| + | |||
| + | The below cycling protocol works for both DREADDs and the iCreER (TRAP2) allele. | ||
| + | # 95.0° for 2 min | ||
| + | # 95.0° for 20 sec | ||
| + | # 65.0° for 15 sec, -0.5° per cycle | ||
| + | # 72.0° for 10 sec | ||
| + | # Goto 2, 10 times | ||
| + | # 95.0° for 15 sec | ||
| + | # 60.0° for 15 sec | ||
| + | # 72.0° for 10 sec | ||
| + | # Goto 6, 30 times | ||
| + | # 72.0° for 5 min | ||
Latest revision as of 12:29, 24 July 2024
Extraction and Purification of DNA from Tail Biopsy
Tail Biopsy
Mice are first anesthetized in a bell jar with Isoflurane for ~20 sec, monitor animal closely and check with toe pinch before proceeding
Cycling Protocol
The below cycling protocol works for both DREADDs and the iCreER (TRAP2) allele.
- 95.0° for 2 min
- 95.0° for 20 sec
- 65.0° for 15 sec, -0.5° per cycle
- 72.0° for 10 sec
- Goto 2, 10 times
- 95.0° for 15 sec
- 60.0° for 15 sec
- 72.0° for 10 sec
- Goto 6, 30 times
- 72.0° for 5 min