PCR-from purified DNA - Revision history

Tkveton: /* Notes */

2015-07-20T19:59:47Z

Notes

← Older revision		Revision as of 19:59, 20 July 2015
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	== Notes ==		== Notes ==
			The varying times for the steps will depend on the conditions for the primer you are using

	== Papers where this or a similar method has been used ==		== Papers where this or a similar method has been used ==

Tkveton: /* Protocol */

2015-07-20T19:59:19Z

Protocol

← Older revision		Revision as of 19:59, 20 July 2015
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	== Protocol ==		== Protocol ==
	-Add to a 0.2 mL PCR tube 10 microliters of LongAmp Taq Reaction Buffer, 1.5 microliters of 10 mM dNTPs, ~~2 microliters~~ of 10 micromolar forward primer, ~~2 microliters~~ of 10 micromolar reverse primer, ~~2 microliters~~ of LongAmpTaq DNA Polymerase and 32.5 microliters of nuclease free water		-Add to a 0.2 mL PCR tube 5 microliters of LongAmp Taq Reaction Buffer, 0.75 microliters of 10 mM dNTPs, 1 microliter of 10 micromolar forward primer, 1 microliter of 10 micromolar reverse primer, 1 microliter of LongAmpTaq DNA Polymerase and 16.25 microliters of nuclease free water

	-Suspend 0.3 microliters of DNA in the tube by mixing with a pipette		-Suspend 0.3 microliters of DNA in the tube by mixing with a pipette

Tkveton: /* Materials Needed */

2015-07-20T19:58:40Z

Materials Needed

← Older revision		Revision as of 19:58, 20 July 2015
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	-Nuclease free water		-Nuclease free water

	-DNA template		-DNA template(purified DNA)

	-PCR machine		-PCR machine

Tkveton: /* Protocol */

2015-07-13T17:54:51Z

Protocol

← Older revision		Revision as of 17:54, 13 July 2015
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	-Add to a 0.2 mL PCR tube 10 microliters of LongAmp Taq Reaction Buffer, 1.5 microliters of 10 mM dNTPs, 2 microliters of 10 micromolar forward primer, 2 microliters of 10 micromolar reverse primer, 2 microliters of LongAmpTaq DNA Polymerase and 32.5 microliters of nuclease free water		-Add to a 0.2 mL PCR tube 10 microliters of LongAmp Taq Reaction Buffer, 1.5 microliters of 10 mM dNTPs, 2 microliters of 10 micromolar forward primer, 2 microliters of 10 micromolar reverse primer, 2 microliters of LongAmpTaq DNA Polymerase and 32.5 microliters of nuclease free water

	-Suspend ~~the~~ DNA in the tube by mixing with a pipette		-Suspend 0.3 microliters of DNA in the tube by mixing with a pipette

	-Set the PCR machine to have an initial denaturation at 94 C for 30 seconds, 30 cycles of 94 C for 10-30 seconds, 45-65 C for 15-60 seconds and 65 C for 50 seconds per kilobase pair of the DNA fragment, then 65 C for 10 minutes for the final extension, and hold at 4-10 C		-Set the PCR machine to have an initial denaturation at 94 C for 30 seconds, 30 cycles of 94 C for 10-30 seconds, 45-65 C for 15-60 seconds and 65 C for 50 seconds per kilobase pair of the DNA fragment, then 65 C for 10 minutes for the final extension, and hold at 4-10 C

Tkveton: /* Materials Needed */

2015-07-09T18:30:45Z

Materials Needed

← Older revision		Revision as of 18:30, 9 July 2015
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	== Materials Needed ==		== Materials Needed ==
			-PCR strip of microtubes

			-LongAmp Taq Reaction Buffer

			-10 mM dNTPs

			-10 micromolar of appropriate forward primer

			-10 micromolar of appropriate reverse primer

			-LongAmpTaq DNA Polymerase

			-Nuclease free water

			-DNA template

			-PCR machine

			-micropipette

	== Protocol ==		== Protocol ==

Tkveton: /* Protocol */

2015-07-09T18:30:38Z

Protocol

← Older revision		Revision as of 18:30, 9 July 2015
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	== Protocol ==		== Protocol ==
	-Add to a 0.2 mL PCR tube 10 microliters of LongAmp Taq Reaction Buffer, 1.5 microliters of 10 ~~minimolar~~ dNTPs, 2 microliters of 10 ~~minimolar~~ forward primer, 2 microliters of 10 ~~minimolar~~ reverse primer, 2 microliters of LongAmpTaq DNA Polymerase and 32.5 microliters of nuclease free water		-Add to a 0.2 mL PCR tube 10 microliters of LongAmp Taq Reaction Buffer, 1.5 microliters of 10 mM dNTPs, 2 microliters of 10 micromolar forward primer, 2 microliters of 10 micromolar reverse primer, 2 microliters of LongAmpTaq DNA Polymerase and 32.5 microliters of nuclease free water

	-Suspend the DNA in the tube by mixing with a pipette		-Suspend the DNA in the tube by mixing with a pipette

Tkveton: /* Protocol */

2015-07-09T18:27:10Z

Protocol

← Older revision		Revision as of 18:27, 9 July 2015
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	== Protocol ==		== Protocol ==
			-Add to a 0.2 mL PCR tube 10 microliters of LongAmp Taq Reaction Buffer, 1.5 microliters of 10 minimolar dNTPs, 2 microliters of 10 minimolar forward primer, 2 microliters of 10 minimolar reverse primer, 2 microliters of LongAmpTaq DNA Polymerase and 32.5 microliters of nuclease free water

			-Suspend the DNA in the tube by mixing with a pipette

			-Set the PCR machine to have an initial denaturation at 94 C for 30 seconds, 30 cycles of 94 C for 10-30 seconds, 45-65 C for 15-60 seconds and 65 C for 50 seconds per kilobase pair of the DNA fragment, then 65 C for 10 minutes for the final extension, and hold at 4-10 C

			-Run the PCR machine with the tube inside

	== Notes ==		== Notes ==

Tkveton: /* Purpose */

2015-07-09T18:26:34Z

Purpose

← Older revision		Revision as of 18:26, 9 July 2015
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	{{Pagetop}}		{{Pagetop}}
	== Purpose ==		== Purpose ==
			This protocol is used to amplify copies of a desired strand of DNA isolated from purified DNA.

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

Tkveton: New Template:Method Page

2015-07-09T18:26:03Z

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]]