Cloning of the mgtc promoter into the pSB1A3 plasmid

Kamil

Tasks:

• Colonies transfer
• Plasmid isolation and digest

Methods:

• Selected white colonies were transfered to a new dish, liquid cultures were established along the way.
• The liquid cultures were incubated for 6h at 37°C with aeration.
• The plasmids were isolated according to the protocol (see at the end of the page).
• The plasmids were digested with EcoRI and PstI enzymes.
• The results were visualised with electrophoresis on 1% agarose gel.

NOTE: Steps 3 to 5 were not carried out by me and I cannot take credit for them.

Results:

• Gel Electrophoresis:

From left:

• GeneRuler DNA Ladder Mix #SM0333 (Fermentas)
• Colonies from 1 to 10
• GeneRuler DNA Ladder Mix #SM0333 (Fermentas)

NOTE: The upper band represents the plasmid backbone, the lower band is a bit odd. The mgtc promoter is roughly 500 bps in length while the only band except the backbone is somewhere around 1500 bps. Ligation picked up something completely wrong.

Conclusions:

• Back to the drawing board, again...

Construction of K177012 operon1_part2

Ania

Tasks:

• Repeating the cloning process again and labeling my samples before going on holidays. Honestly I do not remember now what we did.

• Results:

  Not very promising so far