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Team:Heidelberg/Templates/Del week11 L
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==08-07-2013== | ==08-07-2013== | ||
===Amplification from FS_14 to FS_15; 1.4 kb=== | ===Amplification from FS_14 to FS_15; 1.4 kb=== | ||
[[File:Heidelberg_20130708 no ladder.png|150px|thumb|Analyticals gel older fragments + PCR for amplification of del fragments DelL (08.07), and DelOP (09.07); lane1=Marker, lanes2-7= DelAF (11.4kbp), BB psB4K5 (4.4kbp), DelOP (2.7kbp), DelG (6.4kbp), DelL (1.4kbp), DelAE (5.3kbp), DelEG (11.0kbp), lanes8-10=DelL (1.4kbp), lanes11-13:DelOP (2.5kbp); | [[File:Heidelberg_20130708 no ladder.png|150px|thumb|Analyticals gel older fragments + PCR for amplification of del fragments DelL (08.07), and DelOP (09.07); lane1=Marker, lanes2-7= DelAF (11.4kbp), BB psB4K5 (4.4kbp), DelOP (2.7kbp), DelG (6.4kbp), DelL (1.4kbp), DelAE (5.3kbp), DelEG (11.0kbp), lanes8-10=DelL (1.4kbp), lanes11-13:DelOP (2.5kbp); | ||
run at 100 V, 0.8 % gel (TAE)]] | run at 100 V, 0.8 % gel (TAE)]] | ||
| + | [[File:Heidelberg_2013-07-09-analyt1-7-DelL-DelOPcut.jpg|150px|thumb|Analyticals lane1=Marker, lanes2-7= DelAF (11.4kbp), BB psB4K5 (4.4kbp), DelOP (2.7kbp), DelG (6.4kbp), DelL (1.4kbp), DelAE (5.3kbp), DelEG (11.0kbp) + PCR for amplification of del fragments DelL (08.07), and DelOP (09.07), cut]] | ||
:'''Reaction''' | :'''Reaction''' | ||
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| 1 || 4 || inf | | 1 || 4 || inf | ||
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'''Results:''' | '''Results:''' | ||
Latest revision as of 21:40, 30 September 2013
08-07-2013
Amplification from FS_14 to FS_15; 1.4 kb
- Reaction
| what | µl |
|---|---|
| D. acidovorans DSM-39 | 1 |
| FS_14: (1/10) | 2.5 |
| FS_15: (1/10) | 2.5 |
| Phusion Master Mix | 25 |
| DMSO | 2.5 |
| dd H2O | 16.5 |
- Conditions
| Biorad MyCycler | ||
|---|---|---|
| Cycles-PCR | temperature [°C] | Time [s] |
| 1 | 98 | 10 |
| 12 | 98 | 1 |
| 66 ↓ 0.5 | 5 | |
| 72 | 42 | |
| 18 | 98 | 1 |
| 63 | 5 | |
| 72 | 42 | |
| 1 | 72 | 12 min |
| 1 | 4 | inf |
Results:
- Amplification worked very well, we had bright bands on the right height.
- Bands were cut out and DNA purified using QIAquick Gel Extration Kit.
