Team:TzuChiU Formosa/Notebook/photopaper

From 2011.igem.org

(Difference between revisions)
(2011.09.14-24)
(2011.09.14-24)
Line 234: Line 234:
_______________________________
_______________________________
<br>
<br>
-
total     20μl
+
total      20μl
<gallery>
<gallery>

Revision as of 17:57, 2 October 2011

Contents

Photopaper

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


2011.09.07

'''Rhodobacter rudrum medium'''
K2HPO4 					1g
NaCl					0.5g
FeSO4.7H2O				0.01g
CaCl2 					0.02g
MnCl2.4H2O 				0.002g
MgSO4.7H2O				0.2g
NaMO2O4.2H2O				0.01g
ddH2O					998.258 ml
                                                   (+
______________________________________________________
                                        1L→take100ml
		+
Yeast Extrat				0.5g
Sodium malate
(Sodium succinate dibasic hexohydrate)	5g
NH4Cl 					1g
ddH2O 					893.5ml
                                                  (+
______________________________________________________
                                        1L

'''Raise E. coli(PSB1C3)'''
1.50ml LB+500μl CHLORAMPHENICOL
2.37℃, overnight (14-16hrs)

2011.09.08-13

Plasmid miniprep kit PSB1C3 plasmid

Raise Rhodobacter rubrum

1.50ml LB+500μl CHLORAMPHENICOL 2.37℃, overnight (14-16hrs)

2011.09.09

Raise Gluconacetobacter hansenii 1.50ml LB+500μl CHLORAMPHENICOL 2.37℃, overnight (14-16hrs)

2011.09.10-11

'''Digestion check of DNA''' 
PSB1C3 DNA (control)			3μl

PSB1C3/ EcoR
DNA					500ng
10×buffer				5μl
BSA					5μl
EcoRⅠ					1μl
ddH2O					29μl
                                                  (+
______________________________________________________
                                        50μl

PSB1C3/ pstⅠ
DNA					500ng
10×buffer				5μl
BSA					5μl
pstⅠ					1μl
ddH2O					29μl
                                                  (+
______________________________________________________
                                        50μl

PSB1C3/ EcoRⅠ+pstⅠ
DNA	                                500ng
10×buffer			        5μl
BSA				        5μl
EcoRⅠ					1μl
pstⅠ					1μl
ddH2O					28μl
                                                  (+
______________________________________________________
                                        50μl
→37℃ for 30 mins

'''Digestion of DNA'''
PSB1C3/ EcoRⅠ+pstⅠ
DNA					10μl
10×buffer				5μl
BSA					5μl
EcoRⅠ					1μl
pstⅠ					1μl
ddH2O					28μl
                                                  (+
______________________________________________________
                                        50μl
→37℃ for 2 hrs

'''electroelution Purification'''
PSB1C3 backbone

2011.09.14-24

   PCR
template DNA   1μl
5×Buffer     4μl
2.5μM dNTP    1.6μl
10μM F      1μl
10μM R      1μl
Taq        0.2μl
ddH2O      8.8μl
_______________________________
total      20μl

2011.09.15

'''Genome miniprep'''
Gluconacetobacter hansenii


2011.09.18

'''Gel/PCR DNA extraction'''
Gluconacetobacter hansenii


2011.09.21

'''Digestion of DNA'''
acsAB/ XbaⅠ+SpeⅠ
DNA					10μl
10×buffer				5μl
BSA					5μl
EcoRⅠ					1μl
pstⅠ					1μl
ddH2O					28μl
                                                  (+
______________________________________________________
                                        50μl
→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