Team:UT-Tokyo/Sudoku notebook

From 2010.igem.org

(Difference between revisions)
(Pre-experiment)
(Pre-experiment)
Line 68: Line 68:
2-3, 11 plasmid (7/2 miniprep)
2-3, 11 plasmid (7/2 miniprep)
-
2-3 : 9.5 uL / H / ES (1.5 uL)
+
2-3 : ES
-
11 (1st) : 5.2 uL / H /  ES (1.5 uL) → Judged failure by cut check → miniprep again
+
11 (1st) : ES → Judged failure by cut check → miniprep again
-
11 (2nd) : 20.2 uL / H /  ES (1.5 uL)
+
11 (2nd) : ES
Line 121: Line 121:
11 (7/1 miniprep)
11 (7/1 miniprep)
-
11 : 5.2 uL / H / ES (EcoRI: 1.5 uL, SpeI: 1 uL)
+
11 : ES
Line 138: Line 138:
1, 4 (6/28 enzyme digest → short), 11 plasmid (7/6 miniprep)
1, 4 (6/28 enzyme digest → short), 11 plasmid (7/6 miniprep)
-
1 : 8.5 uL / H / SP (1.5 uL)
+
1 : SP
-
4 : 8.7 uL / M /  EX (1.5 uL)
+
4 : EX
-
234 : 10 uL / M / XP (1.5 uL)
+
234 : XP

Revision as of 02:52, 19 September 2010

Sudoku

Abstract Construct Experiment Notebook Result

Notebook

Pre-experiment

Make construct

7/1

Transformation

2-3 plasmid (6/30 ligation)

< 6/30 transformation → fail >


Inoculation

11 plasmid (6/30 thaw, transformation)


Enzyme digestion

2, 3, 5, 7 plasmid (6/25 miniprep)

2 : 59.3 ng/uL SP

3 : 124.2 ng/uL XP

5 : 81.3 ng/uL SP

7 : 85.4 ng/uL SP


Gel extract

2, 3, 5, 7 plasmid (7/1 enzyme cut)

Label : 100701 数X (2,3,5,9) ゲル抽出


7/2

Miniprep

2-3, 11 plasmid (7/1 inoculation)

2-3 → 105 ng/uL

11 (1st) → 191.3 ng/uL

11 (2nd) → 49.5 ng/uL


Enzyme digestion

2-3, 11 plasmid (7/2 miniprep)

2-3 : ES

11 (1st) : ES → Judged failure by cut check → miniprep again

11 (2nd) : ES


7/3

Gel extract

2-3, 11 plasmid (7/2 enzyme cut)

Label : 100703 X ゲル抽出


7/5

Ligation

2-3 (insert, 7/3 ES enzyme cut) & 4 (vector, probably 6/28 gel extract, EX enzyme cut) / 11 (insert; 7/3 ES enzyme cut) & 7 (vector, probably 6/28 gel extract, EX enzyme cut)

2-3 : 37.5 ng/uL → 5 uL & 4 : 5.2 ng/uL → 5 uL

11 : 4.2 ng/uL → 7.5 uL & 7 : Unknown → 2.5 uL


Transformation

2-3-4 plasmid (7/5 ligation) / 11-7 plasmid (7/5 ligation)


7/6

Inoculation

234 plasmid (7/5 plate)

11-7 plasmid (7/5 plate) → failure


Miniprep

234 plasmid (7/6 plate)


Enzyme digestion

11 (7/1 miniprep)

11 : ES


ethanol precipitation

failure


Inoculation

7 plasmid (6/23 plate)


Enzyme digestion

1, 4 (6/28 enzyme digest → short), 11 plasmid (7/6 miniprep)

1 : SP

4 : EX

234 : XP


Inoculation

5 plasmid (6/23 plate)


Check test

Main construct

Research of SP6 RNApol cDNA and F plasmid

Convert lox71(rev) to lox66(rev)

Make Hin(rev)

Revise MS2 gene

Make location(rev)-rbs(rev)-location(rev)

Copyright © 2010 iGEM UT-Tokyo. All rights reserved.