Week 13
From 2007.igem.org
(Difference between revisions)
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*Ligation for [http://partsregistry.org/Part:BBa_I763031 I763031]. | *Ligation for [http://partsregistry.org/Part:BBa_I763031 I763031]. | ||
*Transformation of [http://partsregistry.org/Part:BBa_I763031 I763031]. | *Transformation of [http://partsregistry.org/Part:BBa_I763031 I763031]. | ||
- | *We inoculate I763036, | + | *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 | + | *Transformation of [http://partsregistry.org/Part:BBa_I763028 I763028]. |
Line 9: | Line 9: | ||
::'''09/25/07''' | ::'''09/25/07''' | ||
*Miniprep of: | *Miniprep of: | ||
- | ::- I763036; | + | ::- [http://partsregistry.org/Part:BBa_I763036 I763036]; |
- | ::- | + | ::- [http://partsregistry.org/Part:BBa_I763007 I763007]; |
::- [http://partsregistry.org/Part:pSB4A3 pSB4A3]; | ::- [http://partsregistry.org/Part:pSB4A3 pSB4A3]; | ||
- | ::- I763033. | + | ::- [http://partsregistry.org/Part:BBa_I763033 I763033]. |
*We inoculate (5ml LB, 5ul amp 1000X) straked transformations of: | *We inoculate (5ml LB, 5ul amp 1000X) straked transformations of: | ||
- | ::- I763033; | + | ::- [http://partsregistry.org/Part:BBa_I763033 I763033]; |
- | ::- | + | ::- [http://partsregistry.org/Part:BBa_I763028 I763028] (vitality test). |
*Transformations of: | *Transformations of: | ||
- | ::-I763031. | + | ::-[http://partsregistry.org/Part:BBa_I763031 I763031]. |
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'''Fluorescence test according to [[Bologna University/Fluorescence Test | fluorescence test protocol]]''' of: | '''Fluorescence test according to [[Bologna University/Fluorescence Test | fluorescence test protocol]]''' of: | ||
- | ::1. '' | + | ::1. ''[http://partsregistry.org/Part:BBa_I763004 I763004]'': |
::::-bacteria use to have the PMT output maximum value; | ::::-bacteria use to have the PMT output maximum value; | ||
::::-normalization of OD value at 0.5; | ::::-normalization of OD value at 0.5; | ||
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::::-thus we decide to grow the OD value to have higher output value. | ::::-thus we decide to grow the OD value to have higher output value. | ||
- | ::2. '' | + | ::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). | + | ::::- 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. | ::::- thus we decide to use lactose. | ||
Line 46: | Line 46: | ||
::'''09/27/07''' | ::'''09/27/07''' | ||
'''Fluorescence test according to [[Bologna University/Fluorescence Test | Fluorescence test protocol]]''' of: | '''Fluorescence test according to [[Bologna University/Fluorescence Test | Fluorescence test protocol]]''' of: | ||
- | ::1. '' | + | ::1. ''[http://partsregistry.org/Part:BBa_I763004 I763004]'': |
::::-output value like to value with OD=0.5. | ::::-output value like to value with OD=0.5. | ||
- | ::2. '' | + | ::2. ''[http://partsregistry.org/Part:BBa_I763028 I763028]'': |
::::-growth in LB medium; | ::::-growth in LB medium; | ||
::::-pellet in M9 with lactose; | ::::-pellet in M9 with lactose; | ||
Line 57: | Line 57: | ||
::::-Output value without IPTG:0.05V; | ::::-Output value without IPTG:0.05V; | ||
::::-Output value with IPTG: about 0V. | ::::-Output value with IPTG: about 0V. | ||
- | ::4. '' | + | ::4. ''[http://partsregistry.org/Part:BBa_I763019 I763019]'': |
::::-Output value without IPTG:0.05V; | ::::-Output value without IPTG:0.05V; | ||
::::-Output value with IPTG: about 0V. | ::::-Output value with IPTG: about 0V. | ||
Line 63: | Line 63: | ||
::-using of low copy plasmid; | ::-using of low copy plasmid; | ||
::-higher OD; | ::-higher OD; | ||
- | ::-I763032 and I763034 fluorescent test. | + | ::-[http://partsregistry.org/Part:BBa_I763032 I763032] and [http://partsregistry.org/Part:BBa_I763034 I763034] fluorescent test. |
- | *We inoculate I763033 and | + | *We inoculate [http://partsregistry.org/Part:BBa_I763033 I763033] and [http://partsregistry.org/Part:BBa_I763028 I763028]. |
::'''09/28/07''' | ::'''09/28/07''' | ||
- | *Digestion of : I763033 with Eco/Spe. | + | *Digestion of : [http://partsregistry.org/Part:BBa_I763033 I763033] with Eco/Spe. |
- | *Ligation with | + | *Ligation with [http://partsregistry.org/Part:BBa_J04431 J04431] (Eco/Xba). |
*We transform and strake on plates. | *We transform and strake on plates. | ||
[[Bologna | Back]] | [[Bologna | Back]] |
Revision as of 10:20, 8 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. 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.