IGEM:Cambridge/2008/Notebook/Turing Pattern Formation/2008/07/25: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
Yee Yen Tang (talk | contribs) No edit summary |
|||
| (14 intermediate revisions by 2 users not shown) | |||
| Line 1: | Line 1: | ||
<hr width="810px"> | |||
<div id="about"> | |||
{{Cambridge08a}} | |||
</div> | |||
<hr width="810px"> | |||
__NOTOC__ | |||
{|{{table}} width="800" | {|{{table}} width="800" | ||
|- | |- | ||
|style="background-color: # | |style="background-color: #444444"|[[Image:Signalling_button.gif|200px|center]] | ||
|style="background-color: #F2F2F2" align="center"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html> </html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}} | |style="background-color: #F2F2F2" align="center"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html> </html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}} | ||
|- | |- | ||
| colspan="2"| | | colspan="2"| | ||
= | =Results from Yesterday= | ||
* Kan resistance of biobricks | * Kan resistance of biobricks | ||
| Line 25: | Line 31: | ||
'''Results''' | |||
Nothing! 1A1 cells were kept in the freedge! B.S. can not be kept in the fridge, low temperatures kill them! | |||
*Test if biobrick of last year are really the one we want | |||
- Yesterday : Single colony of I746001 and I746101 transferred in 10mL of LB + Amp (final concentration 100μg/mL) | |||
- Take 8mL of I746001 (AIP sender) and of I746101 | |||
- Centrifuge them | |||
- Saved 2ml of each in tubes to regrow and add LB | |||
- Miniprep I746001 and I746101 pellets (plasmid mniprep kit) : same protocol but 60μL of Zyppy Glution Buffer in step 8 | |||
- Restriction Digest : use EcoI and Spel restriction enzymes | |||
- Add sequentially : 15μL of SDW + 2μL of 10X Fast Digest Buffer + 2μL of DNA + 1μL of EcoI + 1μL of Spel | |||
- Microfuge tubes | |||
- Incubate for 10min at 37°C | |||
- Heat inactivation of enzymes at 80°C for 5min | |||
- GEL | |||
20μLof samples + 4μL of dye | |||
* Grow 1A1 | |||
- 10mL of LB + 15μL of 1A1 | |||
- Grow at 37°C | |||
<!-- ## Do not edit below this line unless you know what you are doing. ## --> | <!-- ## Do not edit below this line unless you know what you are doing. ## --> | ||
Revision as of 04:35, 5 September 2008
 |
<html><img src="/images/9/94/Report.png" border="0" /></html> Main project page <html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html> </html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html> | ||||||||||||||||||||
Results from Yesterday
Results Nothing! 1A1 cells were kept in the freedge! B.S. can not be kept in the fridge, low temperatures kill them!
- Yesterday : Single colony of I746001 and I746101 transferred in 10mL of LB + Amp (final concentration 100μg/mL) - Take 8mL of I746001 (AIP sender) and of I746101 - Centrifuge them - Saved 2ml of each in tubes to regrow and add LB - Miniprep I746001 and I746101 pellets (plasmid mniprep kit) : same protocol but 60μL of Zyppy Glution Buffer in step 8 - Restriction Digest : use EcoI and Spel restriction enzymes - Add sequentially : 15μL of SDW + 2μL of 10X Fast Digest Buffer + 2μL of DNA + 1μL of EcoI + 1μL of Spel - Microfuge tubes - Incubate for 10min at 37°C - Heat inactivation of enzymes at 80°C for 5min - GEL 20μLof samples + 4μL of dye
- 10mL of LB + 15μL of 1A1 - Grow at 37°C | |||||||||||||||||||||
