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Team:Heidelberg/Templates/Del week12 AE
From 2013.igem.org
(Difference between revisions)
(Created page with "==19-07-2013== ===Amplificaction from FS_02 to FS_03; 5.3 kb=== [[File:20130719 DelRest.png|150px|thumb|+=with DMSO, -=without DMSO; expected amplicon sizes: LP=6.4kb, 8-9=3.1kb...") |
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===Amplificaction from FS_02 to FS_03; 5.3 kb=== | ===Amplificaction from FS_02 to FS_03; 5.3 kb=== | ||
| - | [[File: | + | [[File:Heidelberg_20130719 DelRest.png|150px|thumb|+=with DMSO, -=without DMSO; expected amplicon sizes: LP=6.4kb, 8-9=3.1kb, 21-09 and 20-09=8.1kb, 22-13s=2.6kb, AE=5.3kb; run at 100 V, 0.8 % gel (TAE)]] |
| - | [[File: | + | [[File:Heidelberg_20130719 DelLP DelG 2xDelFG 2xDelOP 2xDelAE cut.png|150px|thumb|Amplifications of 18.07 and 19.07, cut]] |
:'''Reaction''' | :'''Reaction''' | ||
| Line 11: | Line 11: | ||
! what !! µL | ! what !! µL | ||
|- | |- | ||
| - | | ''D. acidovorans'' || 1 | + | | ''D. acidovorans'' DSM-39 || 1 |
|- | |- | ||
| FS_02: (1/10) || 2.5 | | FS_02: (1/10) || 2.5 | ||
| Line 54: | Line 54: | ||
==20-07-2013== | ==20-07-2013== | ||
| - | === | + | ===Amplification from FS_02 to FS_03; 5.3 kb=== |
| - | [[File: | + | [[File:Heidelberg_20130720 M 3xAE OP FS9toFS21besch.png|150px|thumb|Amplification of AE, OP, FG (20.07); expected amplicon sizes: 21-09(FG)=8.1kb, 22-13s(OP)=2.6kb, AE=5.3kb; run at 100 V, 0.8 % gel (TAE)]] |
:'''Reaction''' | :'''Reaction''' | ||
| Line 62: | Line 62: | ||
! what !! µL | ! what !! µL | ||
|- | |- | ||
| - | | ''D. acidovorans'' || 1 | + | | ''D. acidovorans'' DSM-39 || 1 |
|- | |- | ||
| FS_02: (1/10) || 2.5 | | FS_02: (1/10) || 2.5 | ||
Revision as of 01:32, 30 September 2013
Contents |
19-07-2013
Amplificaction from FS_02 to FS_03; 5.3 kb
- Reaction
2x ~50 µL
| what | µL |
|---|---|
| D. acidovorans DSM-39 | 1 |
| FS_02: (1/10) | 2.5 |
| FS_03: (1/10) | 2.5 |
| Phusion flash Master Mix | 25 |
| DMSO | 2.5 |
| dd H2O | 19 |
- Conditions
| Biometra TProfessional Basic | ||
|---|---|---|
| Cycles | temperature [°C] | Time [s] |
| 1 | 98 | 5 |
| 12 | 98 | 1 |
| 66 ↓ 0.5 | 5 | |
| 72 | 2:30 | |
| 18 | 98 | 1 |
| 63 | 5 | |
| 72 | 2:30 | |
| 1 | 72 | 10min |
| 1 | 12 | inf |
Results:
- Amplification of DelAE did not work since a different cycler was used
20-07-2013
Amplification from FS_02 to FS_03; 5.3 kb
- Reaction
| what | µL |
|---|---|
| D. acidovorans DSM-39 | 1 |
| FS_02: (1/10) | 2.5 |
| FS_03: (1/10) | 2.5 |
| Phusion flash Master Mix | 25 |
| DMSO | 2,5 |
| dd H2O | 19 |
- Conditions
| Biorad MyCycler | ||
|---|---|---|
| Cycles | temperature [°C] | Time [s] |
| 1 | 98 | 5 |
| 12 | 98 | 1 |
| 66 ↓ 0.5 | 5 | |
| 72 | 2:30 | |
| 18 | 98 | 1 |
| 63 | 5 | |
| 72 | 2:30 | |
| 1 | 72 | 10min |
| 1 | 12 | inf |
Results:
- Amplification of DelAE worked but a smear occured, therefore bands were cut out carefully and only used for a test restriction digest
