Haynes:Protocols - Revision history

Karmella Haynes: /* Cell Culture: Mammalian */

Karmella Haynes — Tue, 14 May 2013 21:36:17 GMT

Cell Culture: Mammalian

←Older revision		Revision as of 21:36, 14 May 2013
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	* [[Haynes:TransfectionPlasmid_Lipo \| Transfection - Lipofectamine]] - Transfection of plasmid DNA into cells with Lipofectamine		* [[Haynes:TransfectionPlasmid_Lipo \| Transfection - Lipofectamine]] - Transfection of plasmid DNA into cells with Lipofectamine
	* [[Haynes:SplittingCells \| Splitting Cells]] - passaging cells		* [[Haynes:SplittingCells \| Splitting Cells]] - passaging cells
-

	==Protein==		==Protein==

Karmella Haynes — Mon, 13 May 2013 22:15:20 GMT

←Older revision		Revision as of 22:15, 13 May 2013
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	* [[Haynes:MamCultureMedia \| Media formulas]] - cell line-specific formulas		* [[Haynes:MamCultureMedia \| Media formulas]] - cell line-specific formulas
	* [[Haynes:TransfectionPlasmid_Lipo \| Transfection - Lipofectamine]] - Transfection of plasmid DNA into cells with Lipofectamine		* [[Haynes:TransfectionPlasmid_Lipo \| Transfection - Lipofectamine]] - Transfection of plasmid DNA into cells with Lipofectamine
		+	* [[Haynes:SplittingCells \| Splitting Cells]] - passaging cells

Karmella Haynes — Tue, 19 Mar 2013 23:58:08 GMT

Recipes

Karmella Haynes — Tue, 19 Mar 2013 23:12:36 GMT

Recipes

←Older revision		Revision as of 23:12, 19 March 2013
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	* Penicillin/ streptomycin (pen-strep), 5 mL		* Penicillin/ streptomycin (pen-strep), 5 mL
	* [[Haynes:MamCultureMedia \| Appropriate antibiotics]], depending upon formula		* [[Haynes:MamCultureMedia \| Appropriate antibiotics]], depending upon formula
		+
		+	Add FBS and pen-strep directly to the incomplete medium in the original stock bottle. Cap and invert to mix. Filter sterilize using a vacuum filtration unit.
		+	}}

	'''Oligo Annealing Buffer, 10x''' {{hide\|		'''Oligo Annealing Buffer, 10x''' {{hide\|

Karmella Haynes — Tue, 19 Mar 2013 23:10:47 GMT

Recipes

←Older revision		Revision as of 23:10, 19 March 2013
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	* [[Haynes:MamCultureMedia \| Appropriate antibiotics]], depending upon formula		* [[Haynes:MamCultureMedia \| Appropriate antibiotics]], depending upon formula

-	~~Add FBS and pen~~-~~strep directly to the incomplete medium in the original stock bottle~~. ~~Cap and invert to mix. Filter sterilize using a vacuum filtration unit~~.	+	'''Oligo Annealing Buffer, 10x''' {{hide\|
		+	Formula: [1M NaCl; 100 mM Tris-HCl pH 7.4]<br>
		+	Volume: 1000 μL
		+	* 5M NaCl, 200 μL
		+	* 1M Tris-HCl, 10 μL
		+	* dH<sub>2</sub>O, 790 μL
		+
		+	Keep frozen at -20°C.
	}}		}}
		+
		+



	</div>		</div>

Karmella Haynes — Sat, 23 Feb 2013 02:15:15 GMT

Recipes

←Older revision		Revision as of 02:15, 23 February 2013
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	Add dH<sub>2</sub>O directly to the supplier's tube (containing 100 μL of ladder). Add the 20x loading dye. Mix. Aliquot 200 μL portions into five 1.5 mL tubes. Store at -20°C.		Add dH<sub>2</sub>O directly to the supplier's tube (containing 100 μL of ladder). Add the 20x loading dye. Mix. Aliquot 200 μL portions into five 1.5 mL tubes. Store at -20°C.
		+	}}
		+
		+	'''LB (Lennox) Agar plates''' {{hide\|
		+	Volume: 1 L
		+	* Bacto-agar, 15 g
		+	* Caesin tryptone, 10 g
		+	* Yeast extract, 5 g
		+	* NaCl, 5 g
		+	* 1 N NaOH, 1 mL
		+
		+	Dissolve in 1000 mL dH<sub>2</sub>O. Autoclave to sterilize. Cool to ~50°C (warm to touch) before adding antibiotics. Add antibiotic(s) and pour into sterile petri dishes. 1L should yield about 40 plates. Shortcut: Use 25 g Acros LB Broth, Lennox (BP9722-500) granules instead of caesin tryptone, yeast extract, and NaCl.
	}}		}}

Karmella Haynes — Wed, 09 Jan 2013 06:37:47 GMT

DNA Assembly

←Older revision		Revision as of 06:37, 9 January 2013
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	** [[Haynes:Assembly101 \| Model Procedure for Assembling BioBrick Parts: Classic Ligation for Beginners]]		** [[Haynes:Assembly101 \| Model Procedure for Assembling BioBrick Parts: Classic Ligation for Beginners]]
	* [[Haynes:Gibson_Assembly \| Gibson Assembly]]		* [[Haynes:Gibson_Assembly \| Gibson Assembly]]
		+	* [[Haynes:TypeIIS_Assembly \| Type IIS Assembly]] - similar to Golden Gate, Golden Braid, and Moclo

	<br>		<br>

Karmella Haynes — Mon, 07 Jan 2013 18:25:04 GMT

Protocols

←Older revision		Revision as of 18:25, 7 January 2013
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	* [[Haynes:MamCultureMedia \| Media formulas]] - cell line-specific formulas		* [[Haynes:MamCultureMedia \| Media formulas]] - cell line-specific formulas
	* [[Haynes:TransfectionPlasmid_Lipo \| Transfection - Lipofectamine]] - Transfection of plasmid DNA into cells with Lipofectamine		* [[Haynes:TransfectionPlasmid_Lipo \| Transfection - Lipofectamine]] - Transfection of plasmid DNA into cells with Lipofectamine
		+

	==Protein==		==Protein==
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	==Real Time Quantitative PCR==		==Real Time Quantitative PCR==
	* [[Haynes:UPLassay \| Universal Probe Library (UPL) assay]] - for the Roche Light Cycler 480		* [[Haynes:UPLassay \| Universal Probe Library (UPL) assay]] - for the Roche Light Cycler 480
		+

	==RNA & cDNA protocols==		==RNA & cDNA protocols==

Karmella Haynes — Mon, 07 Jan 2013 18:24:39 GMT

Protocols

←Older revision		Revision as of 18:24, 7 January 2013
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	==Real Time Quantitative PCR==		==Real Time Quantitative PCR==
-	* [[Haynes:UPLassay \| Universal Probe Library (UPL) assay]]: for the Roche Light Cycler 480	+	* [[Haynes:UPLassay \| Universal Probe Library (UPL) assay]] - for the Roche Light Cycler 480

		+	==RNA & cDNA protocols==
		+	* [[Haynes:TRIzol_RNeasy \| RNA mini prep - TRIzol/ RNeasy column]]

-	==Resources at OpenWetWare==	+
		+	==Other Resources - OpenWetWare==
	* [http://openwetware.org/wiki/E._coli_genotypes E. coli Strains]		* [http://openwetware.org/wiki/E._coli_genotypes E. coli Strains]
	* [http://openwetware.org/wiki/Materials Materials]: buffer formulas; specific information on commonly used reagents (''e.g.'', LB broth, ampicillin, restriction enzymes, etc.)		* [http://openwetware.org/wiki/Materials Materials]: buffer formulas; specific information on commonly used reagents (''e.g.'', LB broth, ampicillin, restriction enzymes, etc.)

Karmella Haynes — Sat, 17 Nov 2012 02:32:51 GMT

Cell Culture: Mammalian

←Older revision		Revision as of 02:32, 17 November 2012
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	==Cell Culture: Mammalian==		==Cell Culture: Mammalian==
	* [[Haynes:MamCultureMedia \| Media formulas]] - cell line-specific formulas		* [[Haynes:MamCultureMedia \| Media formulas]] - cell line-specific formulas
-	* [[Haynes:~~TransfectionPlasmid~~ \| Transfection]] - Transfection of plasmid DNA into cells	+	* [[Haynes:TransfectionPlasmid_Lipo \| Transfection - Lipofectamine]] - Transfection of plasmid DNA into cells with Lipofectamine

	==Protein==		==Protein==

←Older revision		Revision as of 22:15, 13 May 2013
Line 34:		Line 34:
	* [[Haynes:MamCultureMedia \| Media formulas]] - cell line-specific formulas		* [[Haynes:MamCultureMedia \| Media formulas]] - cell line-specific formulas
	* [[Haynes:TransfectionPlasmid_Lipo \| Transfection - Lipofectamine]] - Transfection of plasmid DNA into cells with Lipofectamine		* [[Haynes:TransfectionPlasmid_Lipo \| Transfection - Lipofectamine]] - Transfection of plasmid DNA into cells with Lipofectamine
		+	* [[Haynes:SplittingCells \| Splitting Cells]] - passaging cells