Team:LMU-Munich/Notebook/Apoptosis

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(Difference between revisions)
(→8-12-2010)
(→8-10-2010)
Line 89: Line 89:
- TEV p14 recogn = 190-6 (Ampicillin)
- TEV p14 recogn = 190-6 (Ampicillin)
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-> Protocol: https://www.openwetware.org/wiki/Transforming_chemically_competent_cells_protocol (3 Transformation)
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-> Protocol: ([[Team:LMU-Munich/Notebook/Protocols/3_Transformation|3 Transformation]])
- We added 2 µl DNA
- We added 2 µl DNA
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- CMV-Promoter Biobrick: BBa_J52034
- CMV-Promoter Biobrick: BBa_J52034
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-> Protocol: http://www.promega.com/protcards/9fb093/fb093.pdf (4 Plasmid extraction from cells)
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-> Protocol:([[Team:LMU-Munich/Notebook/Protocols/4_Plasmid_extraction_from_cells|4 Plasmid extraction from cells]])
- Prepared overnight culture, measured concentration of DNA
- Prepared overnight culture, measured concentration of DNA

Revision as of 12:57, 13 August 2010


Apoptosis Notebook

Contents

Week Days
31 8-02-2010 8-03-2010 8-04-2010 8-05-2010 8-06-2010 8-07-2010 8-08-2010
32 8-09-2010 8-10-2010 8-11-2010 8-12-2010 8-13-2010 8-14-2010 8-15-2010
33 8-16-2010 8-17-2010 8-18-2010 8-19-2010 8-20-2010 8-21-2010 8-22-2010

8-02-2010

  • Some test text here.

8-03-2010

Some test text in bold We created following tests:

  • test1
  • test2
  • test3

8-04-2010

Example of a table

header 1 header 2 header 3
row 1, cell 1 row 1, cell 2 row 1, cell 3
row 2, cell 1 row 2, cell 2 row 2, cell 3

8-05-2010

text

8-06-2010

text

8-07-2010

text

8-08-2010

text

8-09-2010

text

8-10-2010

Transforming competent cells

- eGFP Biobrick: BBa_I714891 SDY_eGFP (Kanamycin)

- TEV recogn N Degron SF3 = pDS7 (Ampicillin)

- TEV p14 recogn = 190-6 (Ampicillin)

-> Protocol: (3 Transformation)

- We added 2 µl DNA

- We plated out 200 µl


Plasmid Isolation

- CMV-Promoter Biobrick: BBa_J52034

-> Protocol:(4 Plasmid extraction from cells)

- Prepared overnight culture, measured concentration of DNA

-> Poor results -> thrown away

8-11-2010

text

8-12-2010

- plasmid extraction of pDS7 (458ng/µl), eGFP (55ng/µl), 190-6 (193ng/µl)

-> Protocol: (4 Plasmid extraktion from cells)

- Restriction digest with EcoRI and PstI in buffer H (for testing DNA is correct)

10µg DNA: pDS7 (2µl), eGFP (15µl), 190-6 (10µl)

-> Protocol: (5 Restriction digest)

- Colony for Plasmidextraktion (CMV, eGFP, pDS7, 190-6) plated

- PhiC31o plated on Ampicillin-Agar, stored at 37°C

- 50% Glycerol made (for the glycerolstock PhiC31o)

8-13-2010

- Agarosegelelectrophoresis with the digestions (CMV, eGFP, pDS7, 190-6), 125V for 30 minutes and then for 20 minutes;

expected DNA bands: 190-6 (4840bp, 1903bp), pDS7 (8027bp, 6bp), CMV (654 bp (Insert), 2079bp (Plasmid)), eGFP (720bp (Insert), 2750bp (Plasmid))

- Correct DNA bands for 190-6 (~4800bp, ~1900bp, ~6700bp (undigested plasmid)) and eGFP (~2000bp (Plasmid), ~750 bp (Insert)); CMV probably not digested (two bands; one probably normal, one supercoiled) and pDS7 not clear

- Restriction digest from CMV (EcoR1, Pst1; 6µl DNA, buffer H) and pDS7 (EcoR1, Spe1; 2µl DNA, buffer B)

Expected DNA bands: CMV see above, pDS7 (3647bp, 3369bp, 1011bp, 6bp)

-> Protocol (5 Restriction digest)

8-14-2010

text

8-15-2010

text

8-16-2010

text

8-17-2010

text

8-18-2010

text

8-19-2010

text

8-20-2010

text

8-21-2010

text

8-22-2010

text

 

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