DIYbio/BOSSlab/Notebook/2011/05/15: Difference between revisions
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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span> | |style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span> | ||
|style="background-color: #F2F2F2" align="center"| | |style="background-color: #F2F2F2" align="center"|[[File:Report.png|frameless|link={{#sub:{{FULLPAGENAME}}|0|-11}}]][[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|[[File:Resultset_previous.png|frameless|link={{#lnpreventry:{{FULLPAGENAME}}}}]][[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]] }}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}} | ||
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* incubating one 1.5 mL tube with remaining transformant (900 uL) at 37 C. no shaking. | * incubating one 1.5 mL tube with remaining transformant (900 uL) at 37 C. no shaking. | ||
* storing the other 1.5 mL tube with remaining transformant (900 uL) at 4 C. | * storing the other 1.5 mL tube with remaining transformant (900 uL) at 4 C. | ||
* incubation started at | * incubation started at 10:57 PM 5/15/2011 | ||
* the incubator is very erratic. I'm trying to keep the temperature at or below 37 C. It does not have a very sensitive control loop. I think it is designed to stay +/- 10 C its set point. It's really more of an oven. | |||
== Ian's writeup == | |||
* E. coli transformation with red fluorescent protein. | |||
* Basic cleanliness techniques - all materials used were edible. We did not eat them :) | |||
* Competent cells, ampicillin, hot plate, shakers, test tubes, autoclave tape, glassware, pressure sterilizer, pipets, Pitri dishes obtained from industry sources and donations. | |||
* Concentration of ampicillin unknown, suspect pre-measured x1000. | |||
* Oven (incubator), refrigerators, meters, bucket, neoprene gloves, bleach (sterilizer) and scales from off-the-shelf and donation sources. | |||
* RFP plasmid obtained from old diybio kit. | |||
* Stepped through protocol thrice before doing real transformation. | |||
* Several times let cells to incubate on ice longer than protocol called for. | |||
* Used pressure sterilizer and propane frier to sterilize agar, glassware. | |||
* Temperature regulation of oven/ incubator a issue - try to keep as close to 37 C w/ o going over. Sometimes as cold as 28 C. | |||
* Made plenty of extra amp lb plates. | |||
Thanks, <br> | |||
- Ian | |||
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Latest revision as of 20:24, 26 September 2017
Project name | Main project page Previous entry |
Photos: http://www.flickr.com/photos/macowell/sets/72157626730414854/ Preflightwhat's our goal: transformation of Invivogen lyocomp cells w/ BBa_J04450 on card (circa 2008). Materials
what are the risks to us
what waste are we gonna generate
mitigation of community risks
Protocolbased on http://www.invivogen.com/PDF/LyoComp_GT116_TDS.pdf
Notes
Ian's writeup
Thanks, |