Transformation: competent DH5⍺ cells with ligation products BBA and BBB
The objective is to transform competent DH5⍺ cells with the ligations products BBA and BBB. Expected Results Some colonies on the petri dishes LB+Cm plated with 50 µL of bacteria transformed with the different ligation products and more on the petri dishes LB+Cm plated with 200 µL of bacteria. Obtained Results We obtained expected results.
The transformation worked. Colonies contain a plasmid with the Chloramphenicol resistance gene, present in pSB1C3. A PCR colonie is necessary to check the size of the plasmid present in colonies, and therefore in order to know if bacteria integrated the correct plasmid. The objective is to make a stock of E.Coli DH5⍺ competent cells for subsequent transformations. NB: The competency of the prepared cells will be tested on the 20/09.
Digestion: PA and PB
Objectives
Transformation: competent DH5⍺ cells with ligation product BBA and BBB
Objectives
Materials
Protocol
Experimental conditions realized :
Transformation protocol:
Results (obtained on the 21/09)
More colonies on the petri dishes plated with the « new » competent bacteria (from 20/09) transformed with the different ligation products.
A bacterial lawn on the LB petri dishes without antibiotic.
No colonies on the LB+Cm petri dish plated with bacteria transformed with no plasmid (- control).Interpretation
Competent cells: E.Coli DH5⍺
Objectives
Materials
Protocol
Competence