http://2013.igem.org/wiki/index.php?title=Team:Heidelberg/Templates/Del_week10_G&feed=atom&action=history
Team:Heidelberg/Templates/Del week10 G - Revision history
2024-03-29T10:12:33Z
Revision history for this page on the wiki
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Nils.kurzawa at 12:29, 30 September 2013
2013-09-30T12:29:25Z
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<td colspan='2' style="background-color: white; color:black;">Revision as of 12:29, 30 September 2013</td>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==05-07-2013==</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>==05-07-2013==</div></td></tr>
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Nils.kurzawa
http://2013.igem.org/wiki/index.php?title=Team:Heidelberg/Templates/Del_week10_G&diff=251464&oldid=prev
Charly: Created page with "==05-07-2013== ===Amplification from FS_08 to FS_11; 6.5 kb=== [[File:Heidelberg_20130705 DelFG DelBB DelFGsmall DelG.png|150px|thumb|Gel electrophoresis of DelF-G, Backbone, D..."
2013-09-30T00:21:30Z
<p>Created page with "==05-07-2013== ===Amplification from FS_08 to FS_11; 6.5 kb=== [[File:Heidelberg_20130705 DelFG DelBB DelFGsmall DelG.png|150px|thumb|Gel electrophoresis of DelF-G, Backbone, D..."</p>
<p><b>New page</b></p><div>==05-07-2013==<br />
<br />
===Amplification from FS_08 to FS_11; 6.5 kb===<br />
<br />
[[File:Heidelberg_20130705 DelFG DelBB DelFGsmall DelG.png|150px|thumb|Gel electrophoresis of DelF-G, Backbone, DelF-G (5.1 kb), DelG (6.4 kb).]]<br />
<br />
[[File:Heidelberg_20130705 DelFG DelBB DelFGsmall DelG cut.png|150px|thumb|after Gel excision]]<br />
<br />
<br />
:'''Reaction'''<br />
{| class="wikitable"<br />
|-<br />
! what !! µl <br />
|-<br />
| ''D. acidovorans'' DSM-39 || 1<br />
|-<br />
| FS_08: (1/10) || 1<br />
|-<br />
| FS_11: (1/10) || 1<br />
|-<br />
| Phusion Master Mix || 10<br />
|-<br />
| dd H<sub>2</sub>O || 6<br />
|-<br />
| DMSO || 1<br />
|}<br />
<br />
:'''Conditions'''<br />
{| class="wikitable"<br />
|-<br />
! colspan="3" | Biorad MyCycler<br />
|-<br />
! Cycles-PCR !! temperature [°C] !! Time [s]<br />
|-<br />
| 1 || 98 || 5<br />
|-<br />
| rowspan="3"| 14 || 98 || 1<br />
|-<br />
| 62 ↓ 0.5 || 5<br />
|-<br />
| 72 || 3:20 min<br />
|-<br />
|rowspan="3"| 16 || 98 || 1<br />
|-<br />
| 60 || 5<br />
|-<br />
| 72 || 3:20 min<br />
|-<br />
| 1 ||72 || 12 min<br />
|-<br />
| 1 || 4 || inf<br />
|}<br />
<br />
'''Results:'''<br />
* A weak band was visible, but it was on the wrong height.<br />
* Accidently the band was cut anyway.<br />
* Either the primers did not bind or the DNA still had to many secondary structures --> the consequence is to change the annealing temperature.<br />
<br />
==07-07-2013==<br />
===Amplification from FS_08 to FS_11; 6.5 kb===<br />
<br />
[[File:Heidelberg_20130708 DelAE DelBB DelG.png|400px|thumb|PCR for amplification of del fragments DelAE (08.07), pSB4K5 (08.07) and DelG (07.07); lane1=Marker, lanes2-4=DelAE (5.3kbp), lanes5-10=pSB4K5 (4.2kbp), lanes11-13:DelG (6.4kbp);<br />
run at 100 V, 0.8 % gel (TAE)]]<br />
<br />
[[File:Heidelberg_20130708 DelAE DelBB DelG cut.png|400px|thumb|PCR for amplification of del fragments DelAE (08.07), pSB4K5 (08.07) and DelG (07.07), cut]]<br />
<br />
:'''Reaction'''<br />
{| class="wikitable"<br />
|-<br />
! what !! µl <br />
|-<br />
| ''D. acidovorans'' DSM-39|| 1<br />
|-<br />
| FS_08: (1/10) || 2.5<br />
|-<br />
| FS_11: (1/10) || 2.5<br />
|-<br />
| Phusion Master Mix || 25<br />
|-<br />
| dd H<sub>2</sub>O || 19<br />
|-<br />
| DMSO || 2.5<br />
|}<br />
<br />
<br />
: '''Conditions '''<br />
{| class="wikitable"<br />
|-<br />
! colspan="3" | Biorad MyCycler<br />
|-<br />
! Cycles-PCR !! temperature [°C] !! Time [s]<br />
|-<br />
| 1 || 98 || 5<br />
|-<br />
| rowspan="3"| 12 || 98 || 1<br />
|-<br />
| 66 ↓ 0.5 || 5<br />
|-<br />
| 72 || 2:30 min<br />
|-<br />
|rowspan="3"| 18 || 98 || 1<br />
|-<br />
| 63 || 5<br />
|-<br />
| 72 || 2:30 min<br />
|-<br />
| 1 ||72 || 10 min<br />
|-<br />
| 1 || 4 || inf<br />
|}<br />
<br />
'''Results:'''<br />
* There was no band visible on the gel.<br />
* Either the primers did not bind or the DNA still had to many secondary structures --> the consequence is to change the annealing temperature.<br />
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Charly