Team:Bologna/Notebook

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--[[User:Giovannimariotta|Giovannimariotta]] 15:43, 23 October 2008 (UTC)
* Digestion and Control Gel Run of the previous amplified constructs :
* Digestion and Control Gel Run of the previous amplified constructs :

Revision as of 15:43, 23 October 2008

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HOME PROJECT TEAM SUBMITTED PARTS MODELING WETLAB LAB-BOOK BIOSAFETY AND PROTOCOLS




Under notebook are all relevant information regarding the work done every week in the laboratory and protocols followed.

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Contents

Protocols

For more information, see Biosafety page
Protocol Biosafety level
Plates preparation
1
Amplification
1
Transformation
1
Inoculation
1
Miniprep
1
Digestion reaction
1
Gel preparation
2
Electrophoretic run
2
Gel extraction
2
Ligation reaction
1
Chemiocompetent cells
1
Mediums and buffers
1
Antibiotics stocks preparation
1
IPTG stocks preparation
1
Fluorescence test
1
M9 supplemented media
1



Week 1: from 07/21/08 to 07/27/08

General Preparations

  1. Preparation of chemiocompetent cells from E. Coli DH5α, Top10 and DB 3.1
  2. Preparation of antibiotic stocks for Ampicillin and Kanamicin
  3. Preparation of LB medium and LB plates for cloning.

Week 2: from 07/28/08 to 08/03/08

  • Eluition and Amplification from 2008 Registry Collection: [http://partsregistry.org/Part:BBa_R0082 R0082,] [http://partsregistry.org/Part:BBa_R0083, R0083], [http://partsregistry.org/wiki/index.php/Part:BBa_M30109 M30109] in TOP10 strain to build and characterize the Light response system to be our spatial selective trigger.
  • Eluition and Amplification from 2008 Registry Collection: [http://partsregistry.org/Part:BBa_E0240 E0240], [http://partsregistry.org/Part:BBa_P1010 pSB3K3_P1010]in DB3.1 and the Practice Promoter Set ([http://partsregistry.org/wiki/index.php?title=Part:BBa_J23103/ J23150, J23151, J23102]) to test and set up the new [http://partsregistry.org/Measurement Biobrick Standard Measurement Protocol]
  • Transformation and Amplification from our Lab Stock of [http://partsregistry.org/Part:BBa_S0100 S0100], [http://partsregistry.org/wiki/index.php?title=Part:BBa_I763005 I763005] and [http://partsregistry.org/Part:BBa_C0051 C0051]
  • Growth Curves of Dh5 Alpha, Top10 and XL1 Blue with Low Medium and High Copy Numbers to assay and define the different kinetics (Further Detail)

Week 3: from 08/04/08 to 08/10/08

--Giovannimariotta 15:43, 23 October 2008 (UTC)

  • Digestion and Control Gel Run of the previous amplified constructs :

1. S0100 E/S
Consistent Part Length
2. PLAC-CI X/P
Consistent Part Length
3. R0083 S/P
Single Vector Band as Expexted. Is Hard to verify the Part length correctness given the small size
4. R0082 S/P
Single Vector Band as Expexted. Is Hard to verify the Part length correctness given the small size
5. C0051 X/P
Consistent Part Length.
7. M30105 E/S
The Part appears not consistent. The Gel has unexpected multiple bands.
8. RBS GFP TAG X/P
Consistent Part Length
9.Pλ GFP X/P
Consistent Part Length.

  • Ligation of R0082 and R0083 with E0240 to obtain a Reporter for the Light Driven Trigger.

Week 4: from 08/11/08 to 08/17/08

Week 5: from 08/18/08 to 08/24/08

Week 6: from 08/25/08 to 08/31/08

Week 7: from 09/01/08 to 09/07/08

Week 8: from 09/08/08 to 09/14/08

Week 9: from 09/15/08 to 09/21/08

Week 10: from 09/22/08 to 09/28/08

Week 11: from 09/29/08 to 10/05/08

Week 12: from 10/06/08 to 10/12/08

Week 13: from 10/13/08 to 10/19/08

Week 14: from 10/20/08 to 10/26/08

Week 15: from 10/27/08 to 10/29/08



















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