Melbourne/Lab Notebook Weeks 9-12

From 2007.igem.org

(Difference between revisions)
(Week 9)
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*Picked 3-4 transformants from "ligation" plates (17/8) and [[Melbourne/Growing up cells|Grew up]] liquid cultures.
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*[[Melbourne/Transformation Protocol|Transformed]] the two ligation reactions in the pink rack in the cold room and plated on Gen + Amp plate.
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*[[Melbourne/Miniprep protocol|Miniprepped]] liquid cultures from 19/8 labeled LA, LB, LC and [[Melbourne/BBa_Q04510|P2 13K]] A,  [[Melbourne/BBa_Q04510|P2 13K]] B.
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*Concentration of DNA was [[Melbourne/DNA concentration measurement|measured]] to be: A- 221ng/ul <BR> B-224ng/ul.<BR>
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*[[Melbourne/Diagnostic Digest|Digested]]:
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*# LA X/S
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*# LB X/S
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*# LC X/S
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*# [[Melbourne/BBa_Q04510|P2 13K]] A X/S
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*# [[Melbourne/BBa_Q04510|P2 13K]] B X/S
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Desired products:
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*# 450bp (95bp different from CI-2 used as control)
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*# 450bp
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*# 450bp
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*# 1kb
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*# 1kb
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*These were [[Melbourne/Loading a DNA gel|run]] on a gel.
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*[[Melbourne/Diagnostic Digest|Digestions]] were set up for ligations.
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<font color=red>OmpR-inverter</font>
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-Vector: OmpR (AmpR) miniprepped [[Melbourne/BBa_R0082|P1 15P]] from 10/7 digested with S/P.
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-Insert: c1 inverter (1kb fragment) [[Melbourne/BBa_Q04510|P2 13K]] A with X/P.
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<font color=red>RBS-LacZ</font>
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-Vector: LacZ (KanR) [[Melbourne/BBa_E0033|P2 9E]] from 6/8 with E/X
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-Insert: RBS (GenR) [[Melbourne/BBa_J61035|P4 8J]] from 6/7 with E/S
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Digested for 2h and heat inactivated for 10min at 80degC.
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*Made [[Melbourne/Making glycerol Stocksl|Glycerol Stocks]] of [[Melbourne/BBa_Q04510|P2 13K]] using dry ice method.
<font size=3><b>22 Aug 2007  
<font size=3><b>22 Aug 2007  

Revision as of 08:25, 9 October 2007

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Contents

Week 9

19 Aug 2007

  • Picked 3-4 transformants from "ligation" plates (17/8) and Grew up liquid cultures.
  • Transformed the two ligation reactions in the pink rack in the cold room and plated on Gen + Amp plate.


20 Aug 2007


Desired products:

    1. 450bp (95bp different from CI-2 used as control)
    2. 450bp
    3. 450bp
    4. 1kb
    5. 1kb
  • These were run on a gel.


21 Aug 2007

OmpR-inverter -Vector: OmpR (AmpR) miniprepped P1 15P from 10/7 digested with S/P. -Insert: c1 inverter (1kb fragment) P2 13K A with X/P.

RBS-LacZ -Vector: LacZ (KanR) P2 9E from 6/8 with E/X -Insert: RBS (GenR) P4 8J from 6/7 with E/S

Digested for 2h and heat inactivated for 10min at 80degC.

22 Aug 2007


23 Aug 2007


24 Aug 2007


25 Aug 2007

Week 10

26 Aug 2007


27 Aug 2007


28 Aug 2007


29 Aug 2007


30 Aug 2007


31 Aug 2007


1 Sept 2007


Week 11

2 Sept 2007


3 Sept 2007


4 Sept 2007


5 Sept 2007


6 Sept 2007


7 Sept 2007


8 Sept 2007


Week 12

9 Sept 2007


10 Sept 2007


11 Sept 2007


12 Sept 2007


13 Sept 2007


14 Sept 2007


15 Sept 2007