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Team:Heidelberg/Templates/Del week10 AE
From 2013.igem.org
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==06-07-2013== | ==06-07-2013== | ||
===Amplification from FS_02 to FS_03; 5.3 kb=== | ===Amplification from FS_02 to FS_03; 5.3 kb=== | ||
| - | [[File: | + | [[File:Heidelberg_20130706 DelEG DelAE.png|100px|thumb|PCR for amplification of del fragments E-G and A-E, AE+=PCR AE with DMSO, AE-=PCR without DMSO, M= Marker; wanted amplicon sizes are: AE=5.3kb, EG=11.0]] |
:'''Reaction''' | :'''Reaction''' | ||
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! what !! µL | ! what !! µL | ||
|- | |- | ||
| - | | ''D. acidovorans'' || 1 | + | | ''D. acidovorans'' DSM-39 || 1 |
|- | |- | ||
| FS_02: (1/10) || 1 | | FS_02: (1/10) || 1 | ||
| Line 20: | Line 21: | ||
| dd H<sub>2</sub>O || 6/7 | | dd H<sub>2</sub>O || 6/7 | ||
|} | |} | ||
| - | [[File: | + | [[File:Heidelberg_20130706 DelEG DelAE cut.png|100px|thumb|PCR for amplification of del fragments AE and EG, cut]] |
:'''Conditions''' | :'''Conditions''' | ||
Latest revision as of 15:21, 21 October 2013
06-07-2013
Amplification from FS_02 to FS_03; 5.3 kb
- Reaction
| what | µL |
|---|---|
| D. acidovorans DSM-39 | 1 |
| FS_02: (1/10) | 1 |
| FS_03: (1/10) | 1 |
| Phusion Master Mix | 10 |
| DMSO | 1/- |
| dd H2O | 6/7 |
- Conditions
| Biorad MyCycler | ||
|---|---|---|
| Cycles-PCR | temperature [°C] | Time [s] |
| 1 | 98 | 5 |
| 12 | 98 | 1 |
| 66 ↓ 0.5 | 5 | |
| 72 | 2:30 min | |
| 18 | 98 | 1 |
| 63 | 5 | |
| 72 | 2:30 min | |
| 1 | 72 | 10 min |
| 1 | 4 | inf |
Results:
- Amplification worked with 5% DMSO
- Repeat Amplification with the same protocol to increase concentration when DNA is extracted from gel slices
