# Title : 03072019-Digestion pattern of pBK549 (Succesful) 🆗

# Date:

03072019

# Objective:

To check if the plasmid extracted from the bacteria that contains that plasmid is what we think it is, according what was sent by Benoit and Agnes from ETH

# Method

  • Digestion with EcoRv and PvuII enzymes from the general stock
  • Using 3ml of bacterial culture from plasmid extraction, using Promega Miniprep Kit

Protocol

Digestion mix

  • Final volume of 20ul
  • 15,5ul MiliQ
  • 2ul plasmid
  • 2ul cut smart buffer
  • 0,25ul of EcoRv
  • 0,25ul of PvuII

Incubation at 37C for 1 hour, without shaking

Imaging


# Results 😊

# Conclusion

  • It is important to use the maximum culture that it says on the protocol for good plasmid extraction. This time Enzo used 3ml , but previusly we were using 10ml , which was overkilling, and it was unsuccesful the digestion. It seems this was the key change to have a right pattern.