Week 13
From 2007.igem.org
(Difference between revisions)
Line 24: | Line 24: | ||
::'''09/26/07''' | ::'''09/26/07''' | ||
- | '''Fluorescence test according to [[Bologna University/Fluorescence | + | '''Fluorescence test according to [[Bologna University/Fluorescence Test1 | fluorescence test protocol]]''' of: |
::1. ''[http://partsregistry.org/Part:BBa_I763004 I763004]'': | ::1. ''[http://partsregistry.org/Part:BBa_I763004 I763004]'': |
Revision as of 09:39, 24 October 2007
- 09/24/07
- Ligation for [http://partsregistry.org/Part:BBa_I763031 I763031].
- Transformation of [http://partsregistry.org/Part:BBa_I763031 I763031].
- We inoculate [http://partsregistry.org/Part:BBa_I763036 I763036], [http://partsregistry.org/Part:BBa_I763007 I763007], [http://partsregistry.org/Part:pSB4A3 pSB4A3] in 5ml of LB.
- Transformation of [http://partsregistry.org/Part:BBa_I763028 I763028].
- 09/25/07
- Miniprep of:
- - [http://partsregistry.org/Part:BBa_I763036 I763036];
- - [http://partsregistry.org/Part:BBa_I763007 I763007];
- - [http://partsregistry.org/Part:pSB4A3 pSB4A3];
- - [http://partsregistry.org/Part:BBa_I763033 I763033].
- We inoculate (5ml LB, 5ul amp 1000X) straked transformations of:
- - [http://partsregistry.org/Part:BBa_I763033 I763033];
- - [http://partsregistry.org/Part:BBa_I763028 I763028] (vitality test).
- Transformations of:
- -[http://partsregistry.org/Part:BBa_I763031 I763031].
- 09/26/07
Fluorescence test according to fluorescence test protocol of:
- 1. [http://partsregistry.org/Part:BBa_I763004 I763004]:
- -bacteria use to have the PMT output maximum value;
- -normalization of OD value at 0.5;
- -output value without IPTG: [0.22, 0.27]V;
- -output value with IPTG:
- -after 10min: [0.18,0.28]V;
- -after 20min: [0.41,0.46]V;
- -after 30min: [0.42,0.70]V;
- -after 60min: [0.48,0.80]V;
- -thus we decide to grow the OD value to have higher output value.
- 1. [http://partsregistry.org/Part:BBa_I763004 I763004]:
- 2. [http://partsregistry.org/Part:BBa_I763028 I763028]
- - we observe bacteria vitality, but there isn’t fluorescence after O/N growth with and without progressive induction with IPTG (1mM, 2mM, 3mM, 10mM every 60min).
- - thus we decide to use lactose.
- 2. [http://partsregistry.org/Part:BBa_I763028 I763028]
- 09/27/07
Fluorescence test according to Fluorescence test protocol of:
- 1. [http://partsregistry.org/Part:BBa_I763004 I763004]:
- -output value like to value with OD=0.5.
- 2. [http://partsregistry.org/Part:BBa_I763028 I763028]:
- -growth in LB medium;
- -pellet in M9 with lactose;
- -we observe them every 30min:
- -bacteria vitality confirmed;
- -bacteria don’t beam fluorescent.
- 3. [http://partsregistry.org/Part:BBa_I763031 I763031]:
- -Output value without IPTG:0.05V;
- -Output value with IPTG: about 0V.
- 4. [http://partsregistry.org/Part:BBa_I763019 I763019]:
- -Output value without IPTG:0.05V;
- -Output value with IPTG: about 0V.
- 1. [http://partsregistry.org/Part:BBa_I763004 I763004]:
Conclusions:
- -using of low copy plasmid;
- -higher OD;
- -[http://partsregistry.org/Part:BBa_I763032 I763032] and [http://partsregistry.org/Part:BBa_I763034 I763034] fluorescent test.
- We inoculate [http://partsregistry.org/Part:BBa_I763033 I763033] and [http://partsregistry.org/Part:BBa_I763028 I763028].
- 09/28/07
- Digestion of : [http://partsregistry.org/Part:BBa_I763033 I763033] with Eco/Spe.
- Ligation with [http://partsregistry.org/Part:BBa_J04431 J04431] (Eco/Xba).
- We transform and strake on plates.