Plasmid Isolation from Bacteria - Revision history

Tkveton: /* Materials Needed */

2015-07-20T20:05:47Z

Materials Needed

← Older revision		Revision as of 20:05, 20 July 2015
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	== Materials Needed ==		== Materials Needed ==
	-Microfuge tubes		-1.5 mL Microfuge tubes

	-Microcolumns		-Microcolumns with 2 mL collection tubes

	-distilled water		-distilled water

Tkveton: /* Protocol */

2015-07-09T19:41:48Z

Protocol

← Older revision		Revision as of 19:41, 9 July 2015
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	-Centrifuge the bacterial sample in a microfuge tube at 15,000 rpm for 30 seconds		-Centrifuge the bacterial sample in a microfuge tube at 15,000 rpm for 30 seconds

	-Dispose of the supernatant and resuspend the pellet in 600 microliters of water		-Dispose of the supernatant and resuspend the pellet in 600 microliters of distilled water

	-Add 100 microliters of Cell Lysis Buffer and mix by shaking		-Add 100 microliters of Cell Lysis Buffer and mix by shaking

Tkveton: /* Materials Needed */

2015-07-09T19:41:37Z

Materials Needed

← Older revision		Revision as of 19:41, 9 July 2015
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	== Materials Needed ==		== Materials Needed ==
			-Microfuge tubes

			-Microcolumns

			-distilled water

			-Cell Lysis Buffer

			-Neutralization Solution (NSC)

			-Endotoxin Removal Wash

			-Column Wash Solution

			-Micropipette

			-Centrifuge

			-Bacteria with plasmid DNA

			-Broth for bacteria to grow in

	== Protocol ==		== Protocol ==

Tkveton: /* Protocol */

2015-07-09T19:39:39Z

Protocol

← Older revision		Revision as of 19:39, 9 July 2015
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	== Protocol ==		== Protocol ==
			-Grow the bacteria to saturation in an appropriate broth media overnight

			-Centrifuge the bacterial sample in a microfuge tube at 15,000 rpm for 30 seconds

			-Dispose of the supernatant and resuspend the pellet in 600 microliters of water

			-Add 100 microliters of Cell Lysis Buffer and mix by shaking

			-Add 350 microliters of NSC and mix by shaking

			-Centrifuge for 3 minutes at 15,000 rpm

			-Transfer the supernatant to a microcolumn and centrifuge for 30 seconds at 15,000 rpm

			-Discard the flow through and wash the column with 200 microliters of Endotoxin Removal Wash, centrifuge for 30 seconds at 15,000 rpm

			-Discard the flow through and wash the column with 400 microliters of Column Wash Solution, centrifuge for 30 seconds at 15,000 rpm

			-Discard the flow through and place the column in a fresh microfuge tube

			-Wash the column with 30-50 microliters of distilled water, depending on how much bacteria was used, let it sit for one minute, then centrifuge for 30 seconds at 15,000 rpm

			-The plasmid is in the water in the microfuge tube

	== Notes ==		== Notes ==

Tkveton: /* Purpose */

2015-07-09T19:31:33Z

Purpose

← Older revision		Revision as of 19:31, 9 July 2015
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	{{Pagetop}}		{{Pagetop}}
	== Purpose ==		== Purpose ==
			This protocol is used to isolate a desired plasmid from a bacteria containing the plasmid.

	== Materials Needed ==		== Materials Needed ==

Tkveton: New Template:Method Page

2015-07-09T19:30:43Z

New Template:Method Page

New page

{{Pagetop}}
== Purpose ==

== Materials Needed ==

== Protocol ==

== Notes ==

== Papers where this or a similar method has been used ==
<PMIDsummary />

== Related Ontology Terms ==

== References ==
{{RefHelp}}
<references/>

[[Category:Methods]]