DIYbio/BOSSlab/Notebook/2011/05/11 - Revision history

Mackenzie L. Cowell at 03:10, 16 May 2011

2011-05-16T03:10:15Z

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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;"> BOSSLAB</span>
	\|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>}}
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Mackenzie L. Cowell: /* Preflight */

2011-05-12T18:58:50Z

Preflight

←Older revision		Revision as of 18:58, 12 May 2011
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	<!-- ##### DO NOT edit above this line unless you know what you are doing. ##### -->		<!-- ##### DO NOT edit above this line unless you know what you are doing. ##### -->
-	==Preflight==	+	== Preflight ==

	=== what's our goal ===		=== what's our goal ===
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	* hot water; combusted propane gas		* hot water; combusted propane gas

-	=== Protocol ===	+	== Protocol ==

	# Thaw competent cells (Lyocomp E. coli) on ice.		# Thaw competent cells (Lyocomp E. coli) on ice.
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	# Incubate plates at 37°C for 18 h.		# Incubate plates at 37°C for 18 h.

		+	== Visual Protocol ==

-		+	<html>
		+	<a href="http://www.flickr.com/photos/macowell/5713410863/" title="2011-05-11 visual protocol by macowell, on Flickr"><img src="http://farm4.static.flickr.com/3421/5713410863_fc62439a0e.jpg" width="500" height="374" alt="2011-05-11 visual protocol"></a>
		+	</html>

	<!-- ##### 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. ##### -->

Mackenzie L. Cowell: /* Entry title */

2011-05-12T18:53:19Z

Entry title

←Older revision		Revision as of 18:53, 12 May 2011
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	<!-- ##### DO NOT edit above this line unless you know what you are doing. ##### -->		<!-- ##### DO NOT edit above this line unless you know what you are doing. ##### -->
-	==~~Entry title~~==	+	==Preflight==
-	* ~~Insert content here~~...	+
		+	=== what's our goal ===
		+	dry-run through transformation protocol w/ BBa_J04450 on card using Invivogen lyocomp cells
		+
		+	* lyocomp 116 cells, e. coli k12 (?), non-pathogenic
		+	* hot water bath; 42C, heat caution
		+	* Ian: "The chemicals we'll be using amount to waste water (E. coli), cheese whey (tryptone), Vegemite (yeast extract & NaCl), tofu firming salts (MgCl2 and MgSO4), jelly (agar) and antibiotics and the concentrations are fairly low, so no special dangers - i.e. don't eat it. Cleanup can be city sewer, since all of these are things that are already found in waste water."
		+
		+	=== what are the risks today to us ===
		+	* trivial; just doing a dry run w/ the water baths
		+
		+	=== what are the risks today to the community ===
		+	* water
		+
		+	=== what waste are we gonna generate ===
		+	* hot water; combusted propane gas
		+
		+	=== Protocol ===
		+
		+	# Thaw competent cells (Lyocomp E. coli) on ice.
		+	# During this time pre-chill sterile centrifuge tubes on ice.
		+	# Gently tap competent cell containers to mix
		+	# Aliquot bacterial suspension in pre-chilled tubes (100 µl per tube).
		+	# To the aliquots add 1 µl of plasmid (RFP PUC19).
		+	# Gently tap centrifuge tubes to mix
		+	# Incubate 30 min on ice.
		+	# During this time pre-heat water bath to 42°C (don’t use heating block).
		+	# Incubate samples at 42°C in the water bath for 60s. Do not shake.
		+	# Immediately chill samples on ice for 2 minutes.
		+	# Add 0.95 ml of room temperature SOC.
		+	# Plate 10 µl of bacterial suspension on antibiotic-containing agar plate. (filter sterilized Kanamycin100 ug/ml?).
		+	# Transfer some bacteria to another antibiotic-containing plate using a loop.
		+	# Incubate plates at 37°C for 18 h.
		+
		+

Mackenzie L. Cowell: Autocreate 2011/05/11 Entry for DIYbio/BOSSlab/Notebook

2011-05-12T18:50:01Z

Autocreate 2011/05/11 Entry for DIYbio/BOSSlab/Notebook

New page

{|{{table}} width="800"
|-
|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span>
|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"|

==Entry title==
* Insert content here...


|}

__NOTOC__
[[category:OWWLabNotebookV1]]