90. Title : 03072019-Digestion pattern of pBK549 (Succesful) :ok:#

90.1. Date:#

03072019

90.2. 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

90.3. 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

90.4. Results 😊#

90.5. 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.