Week 13

From 2007.igem.org

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Line 1: Line 1:
::'''09/24/07'''
::'''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'''
::'''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]. 
Line 10: Line 24:
::'''09/26/07'''
::'''09/26/07'''
-
'''Fluorescence test according to [[Bologna University/Fluorescence issue | fluorescence issue protocol]]'''  of:
+
'''Fluorescence test according to [[Bologna University/Fluorescence Test1 | Fluorescence Test Protocol]]'''  of:
-
1. plac-gfp:
+
::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;
-
- output value without IPTG: [0.22, 0.27]V;
+
::::-output value without IPTG: [0.22, 0.27]V;
-
-output value with IPTG:  
+
::::-output value with IPTG:  
-
                  - after 10min: [0.18,0.28]V;
+
::::::::-after 10min: [0.18,0.28]V;
-
                  -after 20min: [0.41,0.46]V;
+
::::::::-after 20min: [0.41,0.46]V;
-
                  -after 30min: [0.42,0.70]V;
+
::::::::-after 30min: [0.42,0.70]V;
-
                  -after 60min: [0.48,0.80]V;
+
::::::::-after 60min: [0.48,0.80]V;
-
-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. pTetR-LacI-pLac-cI-LacY-GFP
+
::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 31: Line 45:
::'''09/27/07'''
::'''09/27/07'''
-
 
+
'''Fluorescence test according to [[Bologna University/Fluorescence Test1 | 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.
 +
'''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'''
::'''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.
-
 
+
[[Bologna#Diary | Back]]
-
 
+
-
 
+
-
[[Bologna | Back]]
+

Latest revision as of 15:51, 26 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.
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.



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.

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.

Back

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