Melbourne/Lab Notebook Weeks 9-12
From 2007.igem.org
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| + | *Picked 3-4 transformants from "ligation" plates (17/8) and [[Melbourne/Growing up cells|Grew up]] liquid cultures. | ||
| + | *[[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. | ||
| + | *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]]: | ||
| + | *# LA X/S | ||
| + | *# LB X/S | ||
| + | *# LC X/S | ||
| + | *# [[Melbourne/BBa_Q04510|P2 13K]] A X/S | ||
| + | *# [[Melbourne/BBa_Q04510|P2 13K]] B X/S | ||
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| + | Desired products: | ||
| + | *# 450bp (95bp different from CI-2 used as control) | ||
| + | *# 450bp | ||
| + | *# 450bp | ||
| + | *# 1kb | ||
| + | *# 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> | ||
| + | -Vector: OmpR (AmpR) miniprepped [[Melbourne/BBa_R0082|P1 15P]] from 10/7 digested with S/P. | ||
| + | -Insert: c1 inverter (1kb fragment) [[Melbourne/BBa_Q04510|P2 13K]] A with X/P. | ||
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| + | <font color=red>RBS-LacZ</font> | ||
| + | -Vector: LacZ (KanR) [[Melbourne/BBa_E0033|P2 9E]] from 6/8 with E/X | ||
| + | -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
- Miniprepped liquid cultures from 19/8 labeled LA, LB, LC and P2 13K A, P2 13K B.
- Concentration of DNA was measured to be: A- 221ng/ul
B-224ng/ul.
Desired products:
- 450bp (95bp different from CI-2 used as control)
- 450bp
- 450bp
- 1kb
- 1kb
- These were run on a gel.
21 Aug 2007
- Digestions were set up for ligations.
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.
- Made Glycerol Stocks of P2 13K using dry ice method.
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
