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Team:UT-Tokyo/Sudoku lab note
From 2010.igem.org
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[[Image:Sudoku_Assay_flpe.png|200px|thumb|flpe expression check assay]] | [[Image:Sudoku_Assay_flpe.png|200px|thumb|flpe expression check assay]] | ||
The original flpe include two restriction enzyme sites(EcoRI, SpeI). | The original flpe include two restriction enzyme sites(EcoRI, SpeI). | ||
| - | |||
We modified this flpe by PCR: | We modified this flpe by PCR: | ||
| - | 1st | + | 1st -> cloning |
| - | 2nd | + | 2nd -> modified SpeI site |
| - | 3rd | + | 3rd -> modified EcoRI site |
== Convert lox71(rev) to lox66(rev) == | == Convert lox71(rev) to lox66(rev) == | ||
Revision as of 02:54, 19 September 2010
Sudoku
Abstract Construct Lab note Result
Lab note
This is an abstract page. If you can see detail data, please click each protocols.
Main construct
Detail protocols about this is:
June.
July.
Assay
Terminator leak switch
flpe
The original flpe include two restriction enzyme sites(EcoRI, SpeI).
We modified this flpe by PCR:
1st -> cloning
2nd -> modified SpeI site
3rd -> modified EcoRI site
Convert lox71(rev) to lox66(rev)
Convert lox71(rev) to lox66(rev)
Make Hin(rev)
Virus expression check
Anti-RNA repression
Parts list
| Number | Name | Link to BioBrick | Plate coordinate | Vector | Code length |
| 1 | T7 promoter | plate1-6N | pSB1AK8 | 46bp | |
| 2 | rbs | plate1-1H | pSB1A2 | 15bp | |
| 3 | cre recombinase | plate1-5D | pSB1A2 | 1037bp | |
| 4 | double terminator | plate2-24C | pSB1AK3 | 95bp | |
| 5 | lox66 recombinase site | plate1-17J | pSB1A2 | 34bp | |
| 6 | Kan resistance (rev) | plate1-2K | pSB1A2 | 816bp | |
| 7 | rbs (rev) | plate1-1J | pSB1A2 | 15bp | |
| 8 | lox71 recombinase site |
order-made | --- | --- | 34bp |
| 9 | single terminator | plate1-4H | pSB1AK3 | 39bp | |
| 10 | constant express promoter | plate1-18A | pSB1A2 | 35bp | |
| 11 | Tet resistance (rev) | plate1-1N | pSB1A2 | 1191bp | |
| 12 | hixC | plate1-1B | pSB1A2 | 26bp | |
| 13 | --- |
--- | --- | --- | --- |
| 14 | lox66 | plate1-17H | pSB1A2 | 34bp | |
| 15 | rbs-mRFP1-terminator | plate1-22O | pSB1A2 | 861bp | |
| 16 | location sequence(old) |
order | --- | --- | --- |
| 17 | pSP6(rev) |
order | --- | --- | --- |
| 18 | lox2272 |
order | --- | --- | --- |
| 19 | loading sequence(rev) |
order | --- | --- | --- |
| 20 | frt | From HQ | pSB1A2 | 34bp | |
| 21 | hin(true) | From HQ | ??? | 573bp | |
| 22 | hin(rev) |
--- | --- | ??? | 573bp |
| 23 | flpe |
--- | --- | --- | about 1.2kbp |
| 24 | gfp unit |
--- | --- | --- | about 1.2kbp |
| 25 | flpe(rev) |
--- | --- | --- | about 1.2kbp |
