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Team:Aberdeen Scotland/Mini Preps
From 2009.igem.org
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Our mini preps were prepared using QIAprep Spin Miniprep Kits from Qiagen. The protocol is from the handbook, using a Microcentrifuge. | Our mini preps were prepared using QIAprep Spin Miniprep Kits from Qiagen. The protocol is from the handbook, using a Microcentrifuge. | ||
| + | {{:Team:Aberdeen_Scotland/break}} | ||
| + | |||
| + | ==Protocol== | ||
| + | 1. Culture E.coli overnight in 5ml LB medium in a shaking incubator at 37C | ||
| + | 2. Centrifuge culture at 4000 rpm for 10 minutes. | ||
| + | 3. Resuspend pelleted bacterial cells in 250µl Buffer P1 and transfer to a microcentrifuge tube. | ||
| + | 4. Add 250µl Buffer P2 and mix thoroughly by inverting the tube 4-6 times | ||
| + | 5. Add 350µl Buffer N3 and mix immediately and thoroughly by inverting tube 4-6 times. | ||
| + | 6. Centrifuge for 10 min at 13,000 rpm in a table-top microcentrifuge. | ||
| + | 7. Apply supernatant to QIAprep spin column by decanting/pipetting | ||
| + | 8. Centrifuge for 60s at 13,000 rpm and discard flow-through. | ||
| + | 9. Wash column by adding 0.5ml Buffer PB, centrifuge for 60s and discard flow-through. | ||
| + | 10. Wash column by adding 0.75ml Buffer PE, centrifuge for 60s and discard flow-through. | ||
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<table class="nav"> | <table class="nav"> | ||
Revision as of 14:35, 11 August 2009
University of Aberdeen - Pico Plumber
Mini Preps
Our mini preps were prepared using QIAprep Spin Miniprep Kits from Qiagen. The protocol is from the handbook, using a Microcentrifuge.
Protocol
1. Culture E.coli overnight in 5ml LB medium in a shaking incubator at 37C 2. Centrifuge culture at 4000 rpm for 10 minutes. 3. Resuspend pelleted bacterial cells in 250µl Buffer P1 and transfer to a microcentrifuge tube. 4. Add 250µl Buffer P2 and mix thoroughly by inverting the tube 4-6 times 5. Add 350µl Buffer N3 and mix immediately and thoroughly by inverting tube 4-6 times. 6. Centrifuge for 10 min at 13,000 rpm in a table-top microcentrifuge. 7. Apply supernatant to QIAprep spin column by decanting/pipetting 8. Centrifuge for 60s at 13,000 rpm and discard flow-through. 9. Wash column by adding 0.5ml Buffer PB, centrifuge for 60s and discard flow-through. 10. Wash column by adding 0.75ml Buffer PE, centrifuge for 60s and discard flow-through.
 Gel Preperation
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Purification 
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