IGEM:Cambridge/2008/Notebook/Turing Pattern Formation/2008/07/25: Difference between revisions
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{{Cambridge08a}} | |||
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__NOTOC__ | |||
{|{{table}} width="800" | {|{{table}} width="800" | ||
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|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 | ||
{|class="wikitable" style="text-align:center" border="1" | |||
|- | |||
! Biobrick | |||
! Antibiotic | |||
! Contration (μg/mL) | |||
! Observations | |||
! Conclusion | |||
|- | |||
| I746001 || Kan || 25 || Many colonies || Kan Resistance OK | |||
|- | |||
| I746101 || Kan || 25 || Many colonies || Kan Resistance OK | |||
|- | |||
| Control PUC19 || Kan || 25 || No colony || No Kan Resistance | |||
|- | |||
|} | |||
'''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 | |||
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Revision as of 04:35, 5 September 2008
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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 |