Team:UTP-Panama/Week 15

From 2011.igem.org

(Difference between revisions)
(→WET LAB)
(→WET LAB)
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BBa-K381001 part
BBa-K381001 part
Cultivation 9/6/2011
Cultivation 9/6/2011
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1. Appropriately shook the tubes where the cultivations were.
+
1. Appropriately shook the tubes where the cultivations were.<br>
-
2. In each eppendorf 1.5mL of cultivation (9/6/2011) control (+)
+
2. In each eppendorf 1.5mL of cultivation (9/6/2011) control (+)<br>
-
3. In each eppendorf 1.5mL of cultivation (9/6/2011) control (-)
+
3. In each eppendorf 1.5mL of cultivation (9/6/2011) control (-)<br>
-
4. Centrifuged 13000rpm, 1 min
+
4. Centrifuged 13000rpm, 1 min<br>
-
5. Supernatant of eppendorf 6 tubes were eliminated
+
5. Supernatant of eppendorf 6 tubes were eliminated<br>
-
6. Resuspended in 300µL buffer RNasa (P1)
+
6. Resuspended in 300µL buffer RNasa (P1)<br>
-
7. Added 300µL of buffer (P2)  
+
7. Added 300µL of buffer (P2) <br>
-
8. Mixed by inversion
+
8. Mixed by inversion<br>
-
9. It was kept in room temperature during 5 minutes
+
9. It was kept in room temperature during 5 minutes<br>
-
10. Added 300µL of buffer (P3)  
+
10. Added 300µL of buffer (P3) <br>
-
11. Mixed by inversion
+
11. Mixed by inversion<br>
-
12. Kept in ice during 5 minutes
+
12. Kept in ice during 5 minutes<br>
-
13. Centrifuged 13000rpm, 10 min
+
13. Centrifuged 13000rpm, 10 min<br>
-
14. Extracted the supernatant and it was deposited in new eppendorf tubes carefully, avoiding to touch the pellets.
+
14. Extracted the supernatant and it was deposited in new eppendorf tubes carefully, avoiding to touch the pellets.<br>
-
15. Added to each tube 1000µL of PCA (Phenol:Chloroform:Isoamyl alcohol)
+
15. Added to each tube 1000µL of PCA (Phenol:Chloroform:Isoamyl alcohol)<br>
-
16. Mixed by inversion
+
16. Mixed by inversion<br>
-
17. Centrifuged 13000rpm, 5 min
+
17. Centrifuged 13000rpm, 5 min<br>
-
18. Transferred 750µL aqueous phase to new tubes properly labeled.
+
18. Transferred 750µL aqueous phase to new tubes properly labeled.<br>
-
19. Tubes with plasmids and isopropanol were centrifuged 13000rpm, 15 min
+
19. Tubes with plasmids and isopropanol were centrifuged 13000rpm, 15 min<br>
-
20. Then isopropanol was removed
+
20. Then isopropanol was removed<br>
-
21. Added 500µL EtOH 70% to wash it
+
21. Added 500µL EtOH 70% to wash it<br>
-
22. Mixed by inversion
+
22. Mixed by inversion<br>
-
23. Again centrifuged 13000rpm, 5 min
+
23. Again centrifuged 13000rpm, 5 min<br>
-
24. Eliminated EtOH 70% and were dried (air dry technique)
+
24. Eliminated EtOH 70% and were dried (air dry technique)<br>
-
25. Resuspended in 30µL of H20 ultra-pure (it was cold)
+
25. Resuspended in 30µL of H20 ultra-pure (it was cold)<br>
-
26. Tubes were stored in a white box properly labeled  
+
26. Tubes were stored in a white box properly labeled <br>
Note: Volumes of 500µL EtOH 70% and 30µL of H20 ultra-pure refer to that specific amount for each tube.
Note: Volumes of 500µL EtOH 70% and 30µL of H20 ultra-pure refer to that specific amount for each tube.

Revision as of 21:45, 28 September 2011


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Week 15: September 12 to 17

September 12

WET LAB

We grew the BBa-K381001 in liquid LB + chloramphenicol to then continue with DIRTY MINIPREP in order to extract its plasmids.

  1. Took out of the fridge 2 petri dishes with transformed bacteria with BBa-K381001
  2. It was used:
    1. 9.7µL of chloramphenicol
    2. and 10mL of liquid LB
  3. Prepared (+) and (-) controls for each petri dish in falcon tubes
  4. Each falcon tube of (+) control contains:
    1. 9.7µL of chloramphenicol/10mL of Liquid LB
    2. 5mL of liquid LB
    3. Aliquot of the cultivation
  5. The negative controls had almost the same content, excluding chloramphenicol
  6. The 4 falcon tubes were collocated in the incubator until next day


September 13

Scientific Comunication Activities

Presentation (by the students of iGEM UTP-Panama) in the Civil and Electrical Faculties about:
What's SynBio?
Applications of SynBio
What's iGEM?
What are we doing? (our project)
Future projects

WET LAB

Objetives: DIRTY MINI PREP BBa-K381001 part Cultivation 9/6/2011 1. Appropriately shook the tubes where the cultivations were.
2. In each eppendorf 1.5mL of cultivation (9/6/2011) control (+)
3. In each eppendorf 1.5mL of cultivation (9/6/2011) control (-)
4. Centrifuged 13000rpm, 1 min
5. Supernatant of eppendorf 6 tubes were eliminated
6. Resuspended in 300µL buffer RNasa (P1)
7. Added 300µL of buffer (P2)
8. Mixed by inversion
9. It was kept in room temperature during 5 minutes
10. Added 300µL of buffer (P3)
11. Mixed by inversion
12. Kept in ice during 5 minutes
13. Centrifuged 13000rpm, 10 min
14. Extracted the supernatant and it was deposited in new eppendorf tubes carefully, avoiding to touch the pellets.
15. Added to each tube 1000µL of PCA (Phenol:Chloroform:Isoamyl alcohol)
16. Mixed by inversion
17. Centrifuged 13000rpm, 5 min
18. Transferred 750µL aqueous phase to new tubes properly labeled.
19. Tubes with plasmids and isopropanol were centrifuged 13000rpm, 15 min
20. Then isopropanol was removed
21. Added 500µL EtOH 70% to wash it
22. Mixed by inversion
23. Again centrifuged 13000rpm, 5 min
24. Eliminated EtOH 70% and were dried (air dry technique)
25. Resuspended in 30µL of H20 ultra-pure (it was cold)
26. Tubes were stored in a white box properly labeled

Note: Volumes of 500µL EtOH 70% and 30µL of H20 ultra-pure refer to that specific amount for each tube.

September 14

WET LAB

(Objetives or Title): --ESCRIBIR Y MEJORAR LAB--

September 15

WET LAB

(Objetives or Title): --ESCRIBIR Y MEJORAR LAB--

September 16

WET LAB

(Objetives or Title): --ESCRIBIR Y MEJORAR LAB--

September 17

GENERAL SESSION

Morning: Talikng about WET LAB final activities. All the Team (under Human Practice Direction):

Schematization of algorithms and flow chart for a Human Practice Project.
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