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Revision as of 02:00, 11 September 2012 (view source) Ehoyer (Talk | contribs) ← Older edit		Latest revision as of 02:31, 17 September 2012 (view source) DanWinter (Talk | contribs)
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	{{:Team:Macquarie_Australia/Template/MQ12}}		{{:Team:Macquarie_Australia/Template/MQ12}}
-	Plasmid preparation was performed according to the QIAprep MiniPrep Kit by Qiagen. The handbook can be found here.	+	Plasmid preparation was performed according to the QIAprep MiniPrep Kit by Qiagen.
		+
		+
		+	== QIAprep Spin Miniprep Kit Protocol ==
		+
		+	1) Pellet 1-5 mL bacterial culture by centrifugation at >8000 rpm for 3 minutes at room temperature.
		+
		+	2) Resuspend Pelleted bacterial cells in 250 uL Buffer P1.
		+
		+	3) Add 250 uL Buffer P2 and invert tube 4-6 times.
		+
		+	4) Add 350 uL Buffer N3 and invert immediately 4-6 times and centrifuge for 10 minutes at 13,000 rpm.
		+
		+	5) Apply the supernatant to a QIAprep spin column and centrifuge for 30 seconds. Discard the flow-through.
		+
		+	6) Wash the QIAprep spin column with 0.5 mL Buffer PB and centrifuge for 30 seconds. Discard the flow-through.
		+
		+	7) Wash the QIAprep spin column with 0.75 mL Buffer PE and centrifuge for 30 seconds. Discard the flow-through.
		+
		+	8) Centrifuge for 1 minute to remove residual wash buffer.
		+
		+	9) Place the QIAprep spin column into a clean Eppendorf. Elute DNA by adding 50 uL Buffer EB directly to the filter. Tap the column and stand for 1 minute and then centrifuge for 1 minute.

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Latest revision as of 02:31, 17 September 2012

Plasmid preparation was performed according to the QIAprep MiniPrep Kit by Qiagen.

QIAprep Spin Miniprep Kit Protocol

1) Pellet 1-5 mL bacterial culture by centrifugation at >8000 rpm for 3 minutes at room temperature.

2) Resuspend Pelleted bacterial cells in 250 uL Buffer P1.

3) Add 250 uL Buffer P2 and invert tube 4-6 times.

4) Add 350 uL Buffer N3 and invert immediately 4-6 times and centrifuge for 10 minutes at 13,000 rpm.

5) Apply the supernatant to a QIAprep spin column and centrifuge for 30 seconds. Discard the flow-through.

6) Wash the QIAprep spin column with 0.5 mL Buffer PB and centrifuge for 30 seconds. Discard the flow-through.

7) Wash the QIAprep spin column with 0.75 mL Buffer PE and centrifuge for 30 seconds. Discard the flow-through.

8) Centrifuge for 1 minute to remove residual wash buffer.

9) Place the QIAprep spin column into a clean Eppendorf. Elute DNA by adding 50 uL Buffer EB directly to the filter. Tap the column and stand for 1 minute and then centrifuge for 1 minute.