Team:UTP-Panama/Week 11

Week 11: August 15 to 20

August 15

WET LAB SESSION

OBJECTIVE: The results were not expected, perhaps due to the quality of cloranphenicol, for this reason we decided to prepare this antibiotic again. 1. Cloranphenicol preparation. 2. After preparing this antibiotic proceeded to prepare the control samples in 50 mL falcon tubes. 3. We use the concentration of 9.7 L of cloranphenicol by 10 mL of LB liquid. 4. We prepared two samples (control + and control -). 5. In each tube was added 5 mL of LB liquid. This LB containing an appropriate concentration of antibiotic.

August 16

WET LAB

(Wet lab morning session)

OBJECTIVE:: Electrophoresis • Preparation of 1% agarose. • Wet the walls of the chamber. • In a 70 mL Erlenmeyer flask was added agarose and put a minute in the microwave, then take it off shake it and put it in the microwave 20 seconds. Ethidium bromide • Put in the microwave and cooled with tap water to a temperature between 60 ° C and 70 º C. • Put 2.5 to 5 micolitros Ethidium bromide. We mix before pouring it into the camera. • Once the gel solidified cover it with TAE (1X). (A) • Add 35μL of ultra-filtered water the plasmids were at 65.5 ° C. • We eliminate the droplets that were on the walls with the sping-down applied to the tubes. • We put the tubes in the microwave 5 minutes. • We added loading buffer which was frozen thawed How? • Put dots of paper loading buffer. • Of the 35 uL of plasmid take 10 mL (B). • Re mix in the dots and then the wells. • A and B meet. Plasmid preparation and gel electrophoresis. • The plasmids with the loading buffer in the wells

o Note: It is important to place the (-) side of the DNA

• Run the gel for 3 hours. • The plasmids containing épendor labeled with 1, 2, 3 are kept the cooler den -81 º C (they were 6 tubes).

o Note: We make sure that the test is running if you notice bubbles on the side of the electrodes.

August 17

WET LAB

Electrophoresis of the BBa_K381001.

(Wet Lab session : afternoon)

OBJECTIVE: Results of the electrophoresis new transilluminator

1. Place the gel on the transilluminator. 2. We turned on the computer and then ultraviolet light button. 3. Graduate wavelength at 600 nm.

old transilluminator

1. Place the gel in the chamber with the door pisillos. 2. We closed the door and the power button to position 1. 3. White light lit to accommodate the gel in a position to occupy the entire screen. 4. After placing the gel in the correct position and turn off the white light with the camera turned on the UV light. 5. Once we had the gel in position and intensity of light with a better view to stop the gel, we proceeded to take the picture with the smallest team is on the right side of the old transilluminator.

August 18

WET LAB

No LAB

August 19

WET LAB

Inoculation of transformed bacteria to grow them

Materials Falcon tube de 50mL
Solid LB
Stock of chloranphenicol
BBa_K410000, pSB1C3
BBa_K328003, pSB1C3
BBa_K328001, pSB1C3
Six small petri dishes

To fill each petri dish we used the following calculation to obtain the desired concentration:
20mL de LB × (33μg Chloramphenicol)/(mL de LB) × mL/(34mg Chloramphenicol)=19,4μL

We heated solid LB to make it liquid.
We placed it in an incubator to keep it warm.
We placed 20mL of LB in a 50 mL falcon tube and cooled it a bit but avoiding solidification.
We added 19.4 µL of chloramphenicol to the falcon tube.
We poured about 3.5 ml to each Petri dish.
We tooke an aliquot with a handgrip and spread it on a plate with LB and Chloramphenicol.
We repeated the above procedure for each sample containing different BioBrick using a different handgrip to spread.

August 20

Human Practices
We were working on the brochure and other graphic methods for expose the SynBio.

GENERAL SESSION

Organization of Human Practices Project, Planning of the Next weeks in Wet lab.
Modeling strategies for our project.