Team:UTP-Panama/Week 11
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4. After placing the gel in the correct position and turn off the white light with the camera turned on the UV light. | 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. | 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. | ||
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==August 19== | ==August 19== | ||
===WET LAB=== | ===WET LAB=== | ||
- | ( | + | |
- | + | '''Inoculation of transformed bacteria to grow them''' <br> | |
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+ | Materials <br> | ||
+ | Falcon tubes of 50mL <br> | ||
+ | Solid LB <br> | ||
+ | Stock of chloranphenicol <br> | ||
+ | BBa_K410000, pSB1C3 <br> | ||
+ | BBa_K328003, pSB1C3 <br> | ||
+ | BBa_K328001, pSB1C3 <br> | ||
+ | Six small petri dishes <br> | ||
+ | |||
+ | To fill each petri dish we used the following calculation to obtain the desired concentration: <br> | ||
+ | 20mL de LB × (33μg Chloramphenicol)/(mL de LB) × mL/(34mg Chloramphenicol)=19,4μL <br> | ||
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+ | 1. We heated solid LB to make it liquid. <br> | ||
+ | 2. We placed it in an incubator to keep it warm.<br> | ||
+ | 3. We placed 20mL of LB in a 50 mL falcon tube and cooled it a bit but avoiding solidification.<br> | ||
+ | 4. We added 19.4 µL of chloramphenicol to the falcon tube.<br> | ||
+ | 5. We poured about 3.5 ml to each Petri dish.<br> | ||
+ | 6. We tooke an aliquot with a handgrip and spread it on a plate with LB and Chloramphenicol.<br> | ||
+ | 7. We repeated the above procedure for each sample containing different BioBrick using a different handgrip to spread.<br> | ||
==August 20== | ==August 20== |
Latest revision as of 03:59, 29 September 2011
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Week 1 | Week 2 | Week 3 | Week 4 | Week 5 | Week 6 | Week 7 | Week 8 | Week 9 | Week 10 | Week 11 | Week 12 | Week 13 | Week 14 | Week 15 | Week 16 | Week 17 | After Regional Week 1 | After Regional Week 2 | Week 11: August 15 to 20August 15WET LAB SESSIONOBJECTIVE: 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 16WET 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 17WET LABElectrophoresis of the BBa_K381001.
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 19WET LABInoculation of transformed bacteria to grow them Materials To fill each petri dish we used the following calculation to obtain the desired concentration: 1. We heated solid LB to make it liquid. August 20Human Practices
GENERAL SESSIONOrganization of Human Practices Project, Planning of the Next weeks in Wet lab. |