Due to the lack of knowledge and experience in the laboratory, our team did not invent a new system. However, despite this, we still managed to work in the laboratory, and carried out transformations on the cells.

So we came to the following conclusions:
• Plasmid pSB1C3 can be expressed in the bacterial strain Escherichia coli BL21(DE3);
• Chloramphenicol is best diluted in 70% alcohol.

Initially, we used DH5alpha E. coli strains, but the plasmid did not take root in this strain. And we implemented the transformation on E. coli, but with another strain BL21(DE3). Luckily the bacteria have grown! Thus, we understood that the plasmid pSB1C3 that is provided in the BioBricks kit is expressed in Escherichia coli BL21(DE3).

Transformation Protocol:

1. Prepare the required materials:
• BL21(DE3) E.coli competent cells;
• water bath at 42°C;
• ice bucket filled with ice;
• shaking incubator at 37°C;
• alcohol lamp lighter;
• sterile spreading device.

2. Clean the bench with 70% ethanol.

3. Turn on the alcohol lamp.
• Carry out the experiment nearby the alcohol lamp.

4. Take BL21(DE3) E.coli competent cells out of the -80°C freezer and thaw on ice until they are melted.

5. Remove LA agar plate containing antibiotic resistance gene from the refrigerator and warm it at room temperature.

6. Label the LA agar plate (vector name & date).

7. Mix 20 ul of DNA with BL21(DE3) E.coli competent cells in a microcentrifuge tube. Gently mix by flicking the bottom of the tube.

8. Incubate the competent cell and DNA mixture for 30 minutes on ice.

9. Heat shock for 45 sec in a 42°C water bath.

10. Immediately take competent cells out of the water bath.

11. Pour the mixture into the LA plate.

12. Spread the mixture all over LA plate using sterile spreader.

13. Incubate the plate at 37°C overnight.

Chloramphenicol Stock Solution Protocol:

1. Prepare the required materials:
• 0.25-0.5 g of Chloramphenicol;
• 10 ml of 70% EtOH.

2. Weigh 0.25-0.5 g of Chloramphenicol

3. Add 10 ml of 70% EtOH

4. Dissolve completely.

5. Stock may be kept at -20°C for 1 year.