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Contents 1 iGemColombia Notebook 1.1 June 1.1.1 June 30: 1.2 July 1.2.1 July 1 1.2.2 July 5 1.2.3 July 6 1.2.4 July 7 1.2.5 July 8 1.2.6 July 11 1.2.7 July 12 1.2.8 July 14 1.2.9 July 15 1.3 August

iGemColombia Notebook

Here you can find our daily work in the Lab!

June

June 30:

• Biobricks 1, 2, 3, 4 and 5 were resuspended
• Miniprep Solutions (I, II and III) were prepared.

July

July 1

• Biobricks 1, 2, 3, 4 and 5 were resuspended.

July 5

• Biobrick 2 presented no colonies.
• Colonies from bricks 1, 3, 4 and 5 were stinged.

     Task: 
     To make LB medium (15x25mL)

July 6

• Biobricks 1, 3 and 4 were twice plated.
• Brick 5 didn't grow up.
• We've added Ampiciline (3.75mL) and Kanamicine (1.875mL) to the boxed from the previous day.
• LB medium was prepared (15x25mL).
• Liquid LB was prepared (400mL).

     Task: 
     To add ampiciline and tetracicline to the boxes.
     Claim the liquid LB
     Scrape ....
     Sting biobricks 2 and 5 (no clons)
     Pick up the tubes with Merceditas

July 7

• Clons 2 and 5 didn't work out.
• Clons 1, 3 and 4 were planted in LB liquid.
• E. Coli inoculation in Coffee
• Kanamicine resistence plasmid: 0.2 optic density.
• Direct inoculation x 2 and C(-) MgCl2.

     Task: 
     Print electroporation protocol.
     Ask Juan D. Olarte about the inoculation in coffee.
     Minipreps for confirmation of 1, 3 and 4.
     Competent cells for clons 2 and 5.

July 8

• Minipreps for 1, 3 and 4 were made.
• Plant sheets of coffee into different plates: Each sheet was cut in the middle and they were incubated at 25°C and 37°C LB+Kan.

     Task:
     To finish the minipreps from the addition of RNAsa.
     Check the E. Coli growth in coffee.

July 11

• Minipreps have been finished. They've been planted in gel: Low concentration (25 ng/uL).
• Transformation protocol in chemical cells.

     Task:
     Prepare 250 mL of SOC medium
     Print the Transformation protocol for chemical cells.

July 12

• Strains 1 and 4 have been conserved.
• LB liquid culture was prepared again for brick 3.
• E. Coli growth results.

     Task:
     Print the Transformation protocol for chemical cells.
     Competent cells for clons 2 and 5.
     Preserve brick 3.
     Confirm bricks 1, 3 and 4 (Digestion)
     Resuspend all Biobricks.
     Check the E. Coli growth in coffee.

July 14

• E. Coli didn't grow up on the sheets.
• Chemical competent cells were made (DH5α).
• Brick 3 was left to grow in liquid medium.
• Bricks 2, 5, 6,7, 8, 9 and 10 were transformed and plated.

     Task: 
     Print the Transformation protocol for chemical cells.
     Preserve brick 3.
     Confirm all biobricks (Digestion)
     Sting the transformed bricks.
     Add antibiotic to the mediums made today.

July 15

• Brick 3 was preserved in Revco.
• Bricks 2, 5 and 8 were stinged.
• 25 LB+Kan boxes x 25 mL.
• No colonies in brick 6.
• Bricks 7, 9 and 10 were contaminated.

     Task:
     Print the chemical cells protocol.
     Confirm all biobricks (Digestion).
     

August