Team:TzuChiU Formosa/Notebook/photopaper

From 2011.igem.org

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</html>
</html>
 +
<font color="#228B22" size=6>Photopaper</font>
 +
<br><br><font color="#000080" size=4>Meeting Minutes</font>
 +
<Hr Align="left" width="100%" size=2>
 +
<font color="#000000" size=3><b>2011.02.24</b></font>
 +
<br><font size=2><b>Discussion:</b></font>
-
='''Photopaper'''=
 
-
=='''Meeting Notes'''==
 
-
==='''2011.02.24'''===
 
-
'''Discussion:'''
 
*Team organization[[File:001.jpg|right|500px|caption]]
*Team organization[[File:001.jpg|right|500px|caption]]
*Brain storming
*Brain storming
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-
==='''2011.03.04'''===
+
 
-
'''Discussion:'''
+
<br><font color="#000000" size=3><b>2011.03.04</b></font>
 +
<br><font size=2><b>Discussion:</b></font>
 +
 
*Team advisory   
*Team advisory   
*Brain storming
*Brain storming
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-
==='''2011.03.14'''===
+
 
-
'''Discussion:'''
+
 
 +
<br><font color="#000000" size=3><b>2011.03.14</b></font>
 +
<br><font size=2><b>Discussion:</b></font>
 +
 
*Task Allocation
*Task Allocation
*Brain storming
*Brain storming
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-
==='''2011.03.23'''===
+
 
-
'''Discussion:'''
+
<br><font color="#000000" size=3><b>2011.03.23</b></font>
 +
<br><font size=2><b>Discussion:</b></font>
**Project : paperia  
**Project : paperia  
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-
==='''2011.03.24'''===
+
 
-
'''Discussion:'''
+
<br><font color="#000000" size=3><b>2011.03.24</b></font>
 +
<br><font size=2><b>Discussion:</b></font>
* Exp. procedure:  
* Exp. procedure:  
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-
==='''2011.06.22'''===
+
 
-
'''Discussion:'''
+
<br><font color="#000000" size=3><b>2011.06.22</b></font>
 +
<br><font size=2><b>Discussion:</b></font>
 +
 
* Due to some unforseen reason, the team decided to change their project.
* Due to some unforseen reason, the team decided to change their project.
* New project: Biojenny  
* New project: Biojenny  
Line 75: Line 85:
　　　　　　　　　-yeast to be our host
　　　　　　　　　-yeast to be our host
-
==='''2011.07.01'''===
+
 
-
'''Discussion:'''
+
 
 +
<br><font color="#000000" size=3><b>2011.07.01</b></font>
 +
<br><font size=2><b>Discussion:</b></font>
* Freeze > grin > genome DNA isolation > Cloning = silk protein gene
* Freeze > grin > genome DNA isolation > Cloning = silk protein gene
-
==='''2011.07.09'''===
+
 
-
'''Discussion:'''
+
<br><font color="#000000" size=3><b>2011.07.09</b></font>
 +
<br><font size=2><b>Discussion:</b></font>
 +
 
* the connections between 3 silk proteins : Fibl Fibh P25
* the connections between 3 silk proteins : Fibl Fibh P25
* major proteins : H-chain, L-chain, P25
* major proteins : H-chain, L-chain, P25
-
==='''2011.07.15'''===
+
 
-
'''Discussion:'''
+
<br><font color="#000000" size=3><b>2011.07.15</b></font>
 +
<br><font size=2><b>Discussion:</b></font>
 +
 
* Due to limited resources and techniques required, the team decided to switch back to the previous project. Paper making !
* Due to limited resources and techniques required, the team decided to switch back to the previous project. Paper making !
* However it would be modified to be more innovative and creative.
* However it would be modified to be more innovative and creative.
-
==='''2011.07.18'''===
+
 
-
'''Discussion:'''
+
<br><font color="#000000" size=3><b>2011.07.18</b></font>
 +
<br><font size=2><b>Discussion:</b></font>
 +
 
* Latest project : Photo paper
* Latest project : Photo paper
* cyanobacteria is designed as the host, cellulose made up of the glucose produced by cyanobacteria could be one of the main attraction of the project.
* cyanobacteria is designed as the host, cellulose made up of the glucose produced by cyanobacteria could be one of the main attraction of the project.
-
==='''2011.07.23'''===
+
 
-
'''Discussion:'''
+
<br><font color="#000000" size=3><b>2011.07.23</b></font>
 +
<br><font size=2><b>Discussion:</b></font>
* system modification to overcome the problems arises during preliminary round
* system modification to overcome the problems arises during preliminary round
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-
==='''2011.09.15'''===
+
 
 +
 
 +
<br><font color="#000000" size=3><b>2011.09.15</b></font>
'''Genome miniprep'''<br>
'''Genome miniprep'''<br>
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-
==='''2011.09.18'''===
+
 
 +
<br><font color="#000000" size=3><b>2011.09.18</b></font>
'''Gel/PCR DNA extraction'''<br>
'''Gel/PCR DNA extraction'''<br>
Line 120: Line 142:
-
=='''Protocols'''==
+
<font size=4 color="#000080"><b>Protocols</b></font>
 +
<Hr Align="left" width="100%" size=3>
-
==='''2011.09.07'''===
 
-
[[File:Catt.gif|right|250px|caption]]
 
-
'''Rhodobacter rudrum medium'''
 
-
<br>K2HPO4　　　　　　　　　　　　1g
 
-
<br>NaCl　　　　　　　　　　　　　0.5g
 
-
<br>FeSO4．7H2O　　　　　　　　　0.01g
 
-
<br>CaCl2　　　　　　　　　　　　　0.02g
 
-
<br>MnCl2．4H2O　　　　　　　　　0.002g
 
-
<br>MgSO4．7H2O　　　　　　　　　0.2g
 
-
<br>NaMO2O4．2H2O　　　　　　　　0.01g
 
-
<br>ddH2O　　　　　　　　　　　　　998.258 ml
 
-
<br>________________________________________
 
-
<br>　　　　　　　　　　　　　　　　1L　→take100ml
 
-
 
-
　　　　　　　　　　＋　
 
-
 
-
<br>Yeast Extrat　　　　　　　　　　　　　　0.5g
 
-
<br>Sodium malate
 
-
<br>(Sodium succinate dibasic hexohydrate)　　5g
 
-
<br>NH4Cl　　　　　　　　　　　　　　　　　1g
 
-
<br>ddH2O　　　　　　　　　　　　　　　　　893.5ml
 
-
<br>_________________________________________________
 
-
<br>　　　　　　　　　　　　　　　　　　　　1L
 
 +
<br>[[File:1-20110907.jpg|lefe|650px|caption]][[File:Catt.gif|right|250px|caption]]
<br>
<br>
-
'''Raise E. coli(PSB1C3)'''
 
<br>
<br>
-
<br>1.50ml LB+500μl CHLORAMPHENICOL
 
-
<br>2.37℃, overnight (14-16hrs)
 
<br>
<br>
 +
<br>[[File:2-20110908-13.jpg|lefe|650px|caption]]
<br>
<br>
<br>
<br>
-
==='''2011.09.08-13'''===
 
<br>
<br>
-
'''Plasmid miniprep kit'''
+
<br>[[File:3-20110910-11.jpg|lefe|650px|caption]]
-
<br>PSB1C3 plasmid
+
<br>
<br>
<br>
<br>
-
'''Raise Rhodobacter rubrum'''
 
<br>
<br>
-
1.50ml LB+500μl CHLORAMPHENICOL
+
<br>[[File:4-20110912-20.jpg|lefe|650px|caption]]
-
<br>2.37℃, overnight (14-16hrs)
+
<br>
<br>
-
 
-
 
-
==='''2011.09.09'''===
 
-
'''Raise Gluconacetobacter hansenii'''
 
-
<br>1.50ml LB+500μl CHLORAMPHENICOL
 
-
<br>2.37℃, overnight (14-16hrs)
 
<br>
<br>
<br>
<br>
-
 
+
<br>[[File:5-20110921.jpg|lefe|550px|caption]][[File:Cats.jpg|right|404px|caption]]
-
==='''2011.09.10-11'''===
+
<br>
<br>
-
'''Digestion check of DNA'''<br>
 
-
[PSB1C3/EcoRI]
 
<br>
<br>
-
DNA　　　　　　　　　　　500ng<br>
 
-
10×buffer　　　　　　　5μl<br>
 
-
BSA　　　　　　　　　　　5μl<br>
 
-
EcoRⅠ　　　　　　　　　　1μl<br>
 
-
ddH2O　　　　　　　　　29μl<br>
 
-
_______________________________
 
<br>
<br>
-
total　　　　　　　　　　50μl
+
<br>[[File:6-20110922.jpg|lefe|650px|caption]]
<br>
<br>
<br>
<br>
<br>
<br>
-
[PSB1C3/pstⅠ]
+
<br>[[File:7-20110923.jpg|lefe|650px|caption]]
<br>
<br>
-
DNA　　　　　　　　　　　500ng<br>
 
-
10×buffer　　　　　　　　　　5μl<br>
 
-
BSA　　　　　　　　　　　　　5μl<br>
 
-
pstⅠ　　　　　　　　　　　　　1μl<br>
 
-
ddH2O　　　　　　　　　　　29μl<br>                                             
 
-
_________________________________
 
<br>
<br>
-
total　　　　　　　　　　　　50μl
 
-
 
-
[PSB1C3/EcoRⅠ+pstⅠ]
 
<br>
<br>
-
DNA　　　　　　　　　　　　　　500ng<br>
+
<br>[[File:8-20110924.jpg|lefe|550px|caption]][[File:Cats2.gif |right|404px|caption]]
-
10×buffer　　　　　　　　　　　　5μl<br>
+
-
BSA　　　　　　　　　　　　　5μl<br>
+
-
EcoRⅠ　　　　　　　　　　　　1μl<br>
+
-
pstⅠ　　　　　　　　　　　　　1μl<br>
+
-
ddH2O　　　　　　　　　　　　28μl<br>             
+
-
__________________________________
+
-
<br>　　　　　　　　　　　　　　　　
+
-
total　　　　　　　　　　　　　50μl<br>
+
-
→37℃ for 30 mins
+
<br>
<br>
<br>
<br>
<br>
<br>
-
'''Digestion of DNA'''
+
<br>[[File:9-20110925.jpg|lefe|650px|caption]]
<br>
<br>
-
[PSB1C3/EcoRⅠ+pstⅠ]
 
<br>
<br>
-
DNA　　　　　　　　　　　　　　　10μl<br>
 
-
10×buffer　　　　　　　　　　　　5μl<br>
 
-
BSA　　　　　　　　　　　　　　　5μl<br>
 
-
EcoRⅠ　　　　　　　　　　　　　　1μl<br>
 
-
pstⅠ　　　　　　　　　　　　　　　1μl<br>
 
-
ddH2O　　　　　　　　　　　　　　28μl<br>
 
-
____________________________________
 
<br>
<br>
-
total 　　　　　　　　　　　　　50μl<br>
+
<br>[[File:10-20110926.jpg|lefe|650px|caption]]
-
→37℃ for 2 hrs
+
<br>
 +
<br>
 +
<br>
 +
<br>[[File:11-20110927.jpg|lefe|550px|caption]][[File:Cats3.jpg|right|404px|caption]]
 +
<br>
 +
<br>
 +
<br>
 +
<br>[[File:12-20110928.jpg|lefe|650px|caption]]
 +
<br>
 +
<br>
 +
<br>
 +
<br>[[File:13-20110929.jpg|lefe|650px|caption]]
 +
<br>
 +
<br>
 +
<br>
 +
<br>[[File:14-20111001.jpg|lefe|650px|caption]]
<br>
<br>
-
'''electroelution Purification'''<br>
 
-
PSB1C3 backbone
 
-
 
-
==='''2011.09.14-24'''===
 
-
 
-
'''PCR'''<br><br>[[File:Cats2.gif |right|404px|caption]]
 
-
template DNA　　　1μl<br>
 
-
5×Buffer　　　　　4μl<br>
 
-
2.5μM dNTP　　　　1.6μl<br>
 
-
10μM F　　　　　　1μl<br>
 
-
10μM R　　　　　　1μl<br>
 
-
Taq　　　　　　　　0.2μl<br>
 
-
ddH2O　　　　　　8.8μl<br>
 
-
_______________________________
 
<br>
<br>
-
total　　　　　　　20μl
 
-
 
-
 
-
==='''2011.09.21'''===
 
-
'''Digestion of DNA'''
 
-
<br>[acsAB/ XbaⅠ+SpeⅠ]
 
-
<br>DNA　　　　　　　　　　　10μl
 
-
<br>10×buffer　　　　　　　　5μl
 
-
<br>BSA　　　　　　　　　　　5μl
 
-
<br>EcoRⅠ　　　　　　　　　　1μl
 
-
<br>pstⅠ　　　　　　　　　　　1μl
 
-
<br>ddH2O　　　　　　　　　　28μl
 
-
<br>____________________________
 
<br>
<br>
-
total　　　　　　　　　　　　　50μl
 
-
<br>→37℃ for 16 hr
 
-
 
-
'''Digestion of DNA'''
 
-
acsCD/ XbaⅠ+SpeⅠ
 
-
DNA 10μl
 
-
10×buffer 5μl
 
-
BSA 5μl
 
-
EcoRⅠ 1μl
 
-
pstⅠ 1μl
 
-
ddH2O 28μl
 
-
                                                  （＋
 
-
______________________________________________________
 
-
                                        50μl
 
-
→37℃ for 16 hr
 
-
 
-
 
-
 
-
==='''2011.09.22'''===
 
-
 
-
'''Ligation of DNA'''
 
-
PSB1C3-acsAB
 
-
Vector                                  3μl
 
-
Insert                                  14μl
 
-
ligase buffer                          2μl
 
-
ligase                                  1μl
 
-
ddH2O                                  -μl
 
-
                                                  （＋
 
-
______________________________________________________
 
-
                                        20μl
 
-
 
-
'''Ligation of DNA'''
 
-
PSB1A3-acsCD
 
-
Vector                                  3μl
 
-
Insert                                  14μl
 
-
ligase buffer                          2μl
 
-
ligase                                  1μl
 
-
ddH2O                                  -μl
 
-
                                                （＋
 
-
______________________________________________________
 
-
                                        20μl
 
-
 
-
 
-
 
-
==='''2011.09.23'''===
 
-
 
-
'''PCR'''
 
-
PR0011 promoter                        1μl
 
-
5×Buffer 4μl
 
-
2.5μM dNTP 1.6μl
 
-
Taq                         0.2μl
 
-
ddH2O 13.2μl
 
-
                                                  （＋
 
-
______________________________________________________
 
-
                                        20μl
 
-
 
-
 
-
==='''2011.09.24'''===
 
-
 
-
'''Transformation of DNA'''
 
-
PSB1C3-acsAB
 
-
Transform into E.coli
 
-
LB+CHLORAMPHENICOL
 
-
 
-
'''Transformation of DNA'''
 
-
PSB1A3-acsCD
 
-
Transform into E.coli
 
-
LB+Ampicillin
 
-
 
-
'''Transformation of DNA'''
 
-
PSB1C3-acsAB
 
-
PSB1A3-acsCD
 
-
Transform into E.coli
 
-
LB+Ampicillin+CHLORAMPHENICOL
 
-
 
-
 
-
 
-
==='''2011.09.24'''===
 
-
 
-
'''Ligation of DNA'''
 
-
PSB1C3-promoter
 
-
Vector                                  3μl
 
-
Insert                                  14μl
 
-
ligase buffer                          2μl
 
-
ligase                                  1μl
 
-
ddH2O                                  -μl
 
-
                                                  （＋
 
-
______________________________________________________
 
-
                                        20μl
 

Latest revision as of 02:56, 6 October 2011

Photopaper

Meeting Minutes

2011.02.24
Discussion:

  • Team organization
    caption
  • Brain storming
    • paper made by bacteria with add-ons such as colors, fragrance, etc.
    • "light up" the plants for replacing lamp posts.



2011.03.04
Discussion:

  • Team advisory
  • Brain storming
    • Add-ons: colorful cellulose which produced by bacteria such as beta-carotene
    • information exchange with iGEM 2009 Cambridge team




2011.03.14
Discussion:

  • Task Allocation
  • Brain storming
    • Avatar - "light up" the plants by transfecting symbiotic bacteria which cloned into fluorescence gene
    • Eco-friendly warmer - biotic thermal pad



2011.03.23
Discussion:

    • Project : paperia
    • Option 1 : Culture bacteria which has pigment gene
    • Option 2 : Cellulose-producing bacteria secrete pigment into the medium



2011.03.24
Discussion:

  • Exp. procedure:
    • cloning of cellulose gene’s CDS
    • the product should operate within E. coli.



2011.06.22
Discussion:

  • Due to some unforseen reason, the team decided to change their project.
  • New project: Biojenny

　　　　　　　　　-economical and humane way to produce paper in large quantities.
　　　　　　　　　-yeast to be our host



2011.07.01
Discussion:

  • Freeze > grin > genome DNA isolation > Cloning = silk protein gene



2011.07.09
Discussion:

  • the connections between 3 silk proteins : Fibl Fibh P25
  • major proteins : H-chain, L-chain, P25



2011.07.15
Discussion:

  • Due to limited resources and techniques required, the team decided to switch back to the previous project. Paper making !
  • However it would be modified to be more innovative and creative.



2011.07.18
Discussion:

  • Latest project : Photo paper
  • cyanobacteria is designed as the host, cellulose made up of the glucose produced by cyanobacteria could be one of the main attraction of the project.



2011.07.23
Discussion:

  • system modification to overcome the problems arises during preliminary round
  • Biobricks from Tokyo 2010 team will be utilized
    • regulator promoter in order to regulate the secretion of cellulose to solve the aggregation of the bacteria




2011.09.15

Genome miniprep
Gluconacetobacter hansenii



2011.09.18

Gel/PCR DNA extraction
Gluconacetobacter hansenii


Protocols



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Retrieved from "http://2011.igem.org/Team:TzuChiU_Formosa/Notebook/photopaper"