"
Team:Heidelberg/Templates/Del week10 AE
From 2013.igem.org
(Difference between revisions)
(Created page with "==06-07-2013== ===Amplification of FS_02 to FS_03; 5.3 kb=== [[File:20130706 DelEG DelAE.png|100px|thumb|PCR for amplification of del fragments E-G and A-E, AE+=PCR AE with DMSO,...") |
|||
| Line 1: | Line 1: | ||
==06-07-2013== | ==06-07-2013== | ||
| - | ===Amplification | + | ===Amplification from FS_02 to FS_03; 5.3 kb=== |
[[File: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]] | [[File: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]] | ||
Revision as of 22:47, 29 September 2013
06-07-2013
Amplification from FS_02 to FS_03; 5.3 kb
File:20130706 DelEG DelAE.png
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
| what | µL |
|---|---|
| D. acidovorans | 1 |
| FS_02: (1/10) | 1 |
| FS_03: (1/10) | 1 |
| Phusion Master Mix | 10 |
| DMSO | 1/- |
| dd H2O | 6/7 |
File:20130706 DelEG DelAE cut.png
PCR for amplification of del fragments AE and EG, cut
- 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
