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Team:Kyoto/GSDD/Notebook/0921-0927
From 2009.igem.org
- 0727-0802
- Construction.
- 0803-0809
- Construction.
- 0810-0816
- Construction.
- 0817-0823
- Construction.PCR.
- 0824-0830
- Construction.PCR.
- 0831-0906
- Construction.PCR.MPR.
- 0907-0913
- Construction.PCR.MPR.
- 0914-0920
- Construction.PCR.MPR.
- 0921-0927
- Construction.PCR.MPR.
- 0928-1004
- Construction.PCR.MPR.
- 1005-1011
- Construction.PCR.MPR.
- 1012-1018
- Construction.MPR.Observation.
0921-0927 : Construction.PCR.MPR.
Monday, 21 September
To Do
- Colony PCR
- Insert check
- Miniprep
- Restriction enzyme digestion
- Gel extraction and PCR purification
- Ligation
- Transformation
Results
PCR
- ①and ②and ③ are inserted properly.
Miniprep
| sample | conc./(ng/ul) |
|---|---|
| 4(1) | 70.3 |
| 4(2) | 54.8 |
| 4(3)a | 84.7 |
| 1-O-2 | 65.1 |
- Gel extraction
- 4(1)a E-S,1-0-2 X-P
- PCR purification
- 4(2) E-X
Tuesday, 22 September
To Do
- Miniprep 3(1)a1 4(4)a1 11①
- Restriction enzyme digestion
- Gel extraction and PCR purification
- Ligation
- MPR
Results
PCR
Gel extraction and PCR purification
Sample 4(4)E-X, 3(1)E-S
Gel extraction
| sample | conc./(ng/ul) |
|---|---|
| MPR1 | 24.8 |
| MPR2 | 81.0 |
| MPR3 | 60.1 |
| MPR4 | 31.2 |
Wednesday, 23 September
To Do
- Phosphorylation
- Miniprep
- Restriction enzyme digestion
- BAP Gel extraction and PCR purification
- Ligation
Results
electrophoresis
11-1 can be cut by stuI.
4(5) can not be inserted in Byp 561 because 4(5) has PstI site.
Dephosphorylation
- Gel extraction.
| sample | conc./(ng/ul) |
|---|---|
| 4(4)X-P | 22.3 |
| 11-1a BAP | 25.1 |
- PCR purification
| sample | conc./(ng/ul) |
|---|---|
| 4(3)S-P | 134.3 |
| MPR1 | 31.1 |
| MPR2 | 52.2 |
Thursday, 24 September
To Do
- Restriction enzyme digestion (Stu1)
- Transformation
- Transformation
- Miniprep
- Restriction enzyme digestion
- Ligation
- Transformation
- Ligation
- transformation
Results
Miniprep
| sample | conc./(ng/ul) |
|---|---|
| 3(2) | 36.0 |
| 4(5)a | 39.6 |
| 4(5)b | 34.7 |
| 4(5)c | 37.4 |
Friday, 25 September
To Do
- Culture 1M 2M(1) 11 (1)
- Restriction enzyme digestion
- Insert check
- Electorophoresis 4(5)a,b,c
- Colony PCR
Results
Electrophoresis
1M contains product of MPR with 200bp.
2M ,at first, contains product of MPR with 1kbp.
With transformed, it changed to with 600bp.
