| |
| META TOPICPARENT |
name="ProtocolList" |
General Conjugation Protocol |
| |
- Incubate conjugation plate overnight (See image 2 for example of overnight growth).
- Scrape up each conjugation mixture into a micro centrifuge tube with 1 mL PBS. Vortex and gently spin down. Repeat.
- Resuspend conjugation mixture in 1 mL of PBS. Plate 100 µL of this mixture and 100 µL of a 10-fold dilution onto selective plates.
|
|
<
< |
- Pick single colonies and restreak or grown in selective media. Confirm transfer of the plasmid via PCR amplification of the insert.
|
>
> |
- Pick single colonies and restreak or grown in selective media. Confirm transfer of the plasmid via PCR amplification of the insert.
|
| |
Considerations and customization:
Many aspects of this protocol can be tweaked if your initial conjugation attempts are unsuccessful. |
